Transposagen Biopharmaceuticals is a leading company in genome engineering technologies and services. They offer various gene editing tools including NextGEN CRISPR, XTNTM TALENs, and piggyBac transposon system. Their piggyBac system allows for footprint-free gene editing which can precisely edit genes without unwanted changes. They provide custom gene editing reagents and services for applications such as bioproduction, therapeutics, disease models, and more.
This document describes a novel gene targeting approach called aptamer-guided gene targeting (AGT) that uses DNA aptamers selected through capillary electrophoresis systematic evolution of ligands by exponential enrichment to bind site-specific DNA binding proteins and guide donor DNA to specific genetic loci for correction. The approach was shown to increase gene targeting efficiency up to 32-fold in yeast and 16-fold in human cells. It also discusses the potential to develop aptamers for other genome editing tools like CRISPR/Cas9.
Recombinant DNA technology involves isolating and amplifying specific DNA sequences and incorporating them into vector molecules like plasmids or bacterial viruses. This recombinant DNA is then propagated in organisms like E. coli. Genetically engineered bacteria can produce important human proteins like insulin, growth hormone, and clotting factors. Restriction enzymes and DNA ligase are used to cut and join DNA fragments to create recombinant molecules.
Guide to Molecular Cloning - Download the GuideQIAGEN
Molecular cloning can be sometimes tricky with significant challenges involved. Overcome the challenges with the essential knowledge and tips for successful cloning.
Transgene-free CRISPR/Cas9 genome-editing methods in plantsCIAT
"Transgene-free CRISPR/Cas9 genome-editing methods in plants" by Matthew R. Willmann, Ph.D. Director, Plant Transformation Facility College of Agriculture and Life Sciences, School of Integrative Plant Science, Cornell University.
The document discusses genome assembly and finishing processes. It begins by outlining typical project goals of completely restoring the genome and producing a high-quality consensus sequence. It then describes the evolution of sequencing technologies from Sanger to newer platforms and their impact on draft assemblies. Key steps in the assembly and finishing process include library preparation, assembly, identifying gaps, and improving consensus quality.
GENESIS™: Comprehensive genome editing - Translating genetic information into personalised medicines.
Horizon is the only source of rAAV expertise and is uniquely capable of exploiting multiple platforms: CRISPR, ZFNs and rAAV singularly or combined. Horizon’s scientists are experts at all forms of gene editing and so have the experience to help guide customers towards the approach that best suits their project
In addition to a standard gene synthesis service, IDT offers a novel, rapid, and reliable method to build and clone the genes you need at a fraction of the cost of full gene synthesis services. gBlocks® Gene Fragments are double-stranded, sequence-verified DNA blocks of length 125–750 bp. Their high sequence fidelity and rapid delivery time make gBlocks Gene Fragments ideal for a large range of synthetic biology applications. In this presentation, Dr Adam Clore reviews a variety of uses of gBlocks fragments, including CRISPR-based genome modification, qPCR and HRM controls, and the assembly of gene fragments using the Gibson Assembly® Method.
This document describes a novel gene targeting approach called aptamer-guided gene targeting (AGT) that uses DNA aptamers selected through capillary electrophoresis systematic evolution of ligands by exponential enrichment to bind site-specific DNA binding proteins and guide donor DNA to specific genetic loci for correction. The approach was shown to increase gene targeting efficiency up to 32-fold in yeast and 16-fold in human cells. It also discusses the potential to develop aptamers for other genome editing tools like CRISPR/Cas9.
Recombinant DNA technology involves isolating and amplifying specific DNA sequences and incorporating them into vector molecules like plasmids or bacterial viruses. This recombinant DNA is then propagated in organisms like E. coli. Genetically engineered bacteria can produce important human proteins like insulin, growth hormone, and clotting factors. Restriction enzymes and DNA ligase are used to cut and join DNA fragments to create recombinant molecules.
Guide to Molecular Cloning - Download the GuideQIAGEN
Molecular cloning can be sometimes tricky with significant challenges involved. Overcome the challenges with the essential knowledge and tips for successful cloning.
Transgene-free CRISPR/Cas9 genome-editing methods in plantsCIAT
"Transgene-free CRISPR/Cas9 genome-editing methods in plants" by Matthew R. Willmann, Ph.D. Director, Plant Transformation Facility College of Agriculture and Life Sciences, School of Integrative Plant Science, Cornell University.
The document discusses genome assembly and finishing processes. It begins by outlining typical project goals of completely restoring the genome and producing a high-quality consensus sequence. It then describes the evolution of sequencing technologies from Sanger to newer platforms and their impact on draft assemblies. Key steps in the assembly and finishing process include library preparation, assembly, identifying gaps, and improving consensus quality.
GENESIS™: Comprehensive genome editing - Translating genetic information into personalised medicines.
Horizon is the only source of rAAV expertise and is uniquely capable of exploiting multiple platforms: CRISPR, ZFNs and rAAV singularly or combined. Horizon’s scientists are experts at all forms of gene editing and so have the experience to help guide customers towards the approach that best suits their project
In addition to a standard gene synthesis service, IDT offers a novel, rapid, and reliable method to build and clone the genes you need at a fraction of the cost of full gene synthesis services. gBlocks® Gene Fragments are double-stranded, sequence-verified DNA blocks of length 125–750 bp. Their high sequence fidelity and rapid delivery time make gBlocks Gene Fragments ideal for a large range of synthetic biology applications. In this presentation, Dr Adam Clore reviews a variety of uses of gBlocks fragments, including CRISPR-based genome modification, qPCR and HRM controls, and the assembly of gene fragments using the Gibson Assembly® Method.
The CRISPR/Cas9 system has emerged as one of the leading tools for modifying genomes of organisms ranging from E. coli to humans. Additionally, the simple gene targeting mechanism of CRISPR technology has been modified and adapted to other applications that include gene regulation, detection of intercellular trafficking, and pathogen detection. With a wealth of methods for introducing Cas9 and gRNAs into cells, it can be challenging to decide where to start. In this presentation, Dr Adam Clore describes the CRISPR mechanism and some of the most prominent uses for CRISPR, along with methods where IDT technologies can assist scientists in designing, testing, and executing a variety of CRISPR-mediated experiments. For more informaton, visit: http://www.idtdna.com/crispr
IDT provides oligonucleotides and panels for targeted sequencing including stocked and custom gene panels. Their panels include 264 genes for acute myeloid leukemia, 127 genes for pan-cancer analysis, and 4503 genes for inherited diseases. IDT probes are individually synthesized and quality controlled before being pooled. Universal blockers improve on-target rates by blocking adapter participation. Additional services include custom barcoded adapters and gBlocks fragments for quality control.
Targeted Breeding Applications of CRISPR-CasKate Barlow
Doane Chilcoat, Director, Applied Technology Systems, DuPont Pioneer
CRISPR-Cas as an advanced plant breeding tool is a more efficient way to improve plants and help farmers produce more and better food, with fewer resources. The superior properties of CRISPR-Cas allows DuPont Pioneer scientists to develop innovative and sustainable seed products for growers similar to those realized through conventional plant breeding, but with even greater efficiency, accuracy and quality. Pioneer is leading the application of this tool to develop customized agriculture solutions. In this talk, potential product targets of this promising technology will be discussed. Approaches to fostering social license and developing an open innovation model for CRISPR-Cas will also be reviewed.
Genome engineering using CRISPR/Cas9 has several advantages over traditional gene targeting methods: it is faster, more precise, applicable to many species, and less expensive. CRISPR/Cas9 uses the Cas9 nuclease guided by a single guide RNA to introduce double-strand breaks at targeted genomic loci. This can generate gene knockouts through error-prone non-homologous end joining or allow for targeted insertions and modifications through homology-directed repair. While CRISPR/Cas9 has great potential, careful design of guide RNAs and donor templates is needed to minimize off-target effects.
Recent breakthroughs in genome editing technology have led to a rapid adoption that parallels that seen with RNAi. And like RNAi, these methods are taking the scientific world by storm, with high profile publications in fields as diverse as HIV treatment, stem cell therapy, food crop modification and drug development to name but a few.
Critically, the endogenous modification of genes enables the study of their function in a physiological context. It also overcomes some of the artefacts that can result from established techniques such as transgenesis and RNAi, which have mislead researchers with false positives or negatives. Until recently however genome editing required considerable technical expertise, and consequently was a relatively niche pursuit.
In this talk we will look at how the latest developments in genome editing tools have changed this, with improvements in both ease-of-use and targeting efficiency, as well as a concomitant reduction in costs opening up these approaches to the wider scientific community.
Rapid adoption of the CRISPR/Cas9 system has for example led to a long list of organisms and tissues in which genetic changes have been made with high efficiency. Other technologies such as recombinant adeno-associated virus (rAAV) offer further precision, stimulating the cell’s high-fidelity DNA repair pathways to insert exogenous sequence with unrivalled specificity. Targeting efficiency can be improved still further by using the technologies in combination – genome cutting induced by CRISPR can significantly enhance homologous recombination mediated by rAAV.
Despite these rapid advances, some pitfalls remain, and so we’ll discuss some of the key considerations for avoiding these, ranging from simply picking the right tool for the job to designing an experiment that maximises chances of success.
Finally we’ll look at how genome editing is being applied to both basic and translational research, and in both a gene-specific and genome wide manner. For the study of disease associated genes and mutations scientists can now complement wide panels of tumour cells with genetically defined isogenic cell pairs identical in all but precise modifications in their gene of interest. The ease-of-design and efficiency of the CRISPR system is also being exploited for genome wide synthetic lethality screens, facilitating rapid drug target identification with significantly reduced risk of false negatives and off-target false positives. And again, further synergies are achieved when these approaches are combined to look for potential synthetic lethal targets in specific genomic contexts.
The document summarizes molecular characterization of Puccinia striiformis f.sp. tritici (Pst) isolates from Western Canada. Pst isolates were sequenced using Illumina platforms and assembled de novo. Phylogenetic trees were constructed based on rRNA sequences and whole genome assemblies. Comparisons between old isolates from 1990-1993 and new isolates from 2007-2012 identified unique and enriched gene sequences, suggesting genome reorganization in Pst. Functional annotation revealed differences in biological processes between old and new isolates, such as transport and response to exogenous molecules in new isolates.
Fast and Efficient Post-Bisulfite-Seq Library Construction with QIAseq Ultral...QIAGEN
The document describes a new method called post-bisulfite next-generation sequencing library construction (PBLC) for whole genome bisulfite sequencing (WGBS) that overcomes challenges with traditional WGBS workflows. PBLC involves bisulfite treatment before library preparation, which fragments the DNA so no additional fragmentation is needed. This reduces workflow time and enables epigenomic studies from lower DNA inputs. Comparison studies show PBLC produces higher library yields than traditional WGBS and comparable sequencing results, making it a more efficient and sensitive method for WGBS.
This document provides an overview of CRISPR/Cas9 genome editing. It discusses the history and limitations of prior genome engineering techniques like recombinant DNA and zinc finger nucleases. It then explains how CRISPR/Cas9 works as a RNA-guided DNA endonuclease and how this allows it to efficiently and specifically edit genomes. The document outlines several applications of CRISPR/Cas9 like generating knockout animals and cell lines. It also notes some concerns about using the technique for human genome editing.
This document discusses genome editing using the CRISPR-Cas9 system. It begins by introducing three main genome editing technologies - zinc-finger nucleases, TALENs, and the CRISPR-Cas9 system. It then describes the key events in the discovery of CRISPR-Cas9, including its origins as a bacterial defense system. The document outlines the main components of the CRISPR-Cas9 system, including crRNA, tracrRNA, sgRNA, and Cas9. It also summarizes the two main steps in genome editing using CRISPR-Cas9 - knocking out genes and DNA repair. The document concludes by discussing opportunities for applying CRISPR-Cas9 technology across various
The CRISPR/Cas9 system has emerged as one of the leading tools for modifying the genomes of organisms ranging from E. coli to humans. In this presentation, we discuss various methods for generating the crRNA and tracrRNA components that are required for guiding the Cas9 endonuclease to genomic targets. You will also learn how to optimize a new 2-part CRISPR RNA system from IDT that offers multiple benefits over other technologies.
Multi Target Gene Editing using CRISPR Technology for Crop ImprovementTushar Gajare
This document provides an overview of a presentation on using CRISPR technology for multi-target gene editing in crop improvement. It begins with an introduction to genome editing and CRISPR-Cas9. It then discusses the CRISPR system, how CRISPR-Cas9 works, its history and applications for crop improvement including case studies in maize with high mutant efficiencies and targeted mutagenesis of multiple genes. The presentation covers advantages and limitations of the technology as well as future prospects.
The document provides an overview of gene knockout and homology-directed repair using CRISPR. It discusses designing guide RNAs and comparing delivery methods like lipofection, electroporation, and microinjection. It also covers designing repair templates for homology-directed repair to insert or change DNA sequences. Optimization of guide RNAs, delivery method, and repair template design can improve genome editing efficiency.
This document summarizes QIAGEN's products for sample preparation, targeted sequencing, and single-cell analysis across various areas of biomedical research including liquid biopsy, circulating tumor cells, and gene expression profiling. Key products mentioned are the QIAseq cfDNA All-in-One Kits for streamlined library preparation from plasma/serum, QIAseq Targeted DNA/RNA Panels for digital sequencing of genomic regions or genes, and QIAseq FX Single Cell DNA/RNA Library Kits for cell-to-library workflows from isolated single cells.
Ernesto Picardi – Bioinformatica e genomica comparata: nuove strategie sperim...eventi-ITBbari
Bioinformatica e genomica comparata: nuove strategie sperimentali e computazionali per la produzione e analisi di dati NGS finalizzati a sviluppare processi e prodotti innovativi per la salute dell’uomo, l’ambiente e l’agroalimentare.
CRISPR-Cas9 is a powerful tool for genome engineering. The document provides guidance on using CRISPR-Cas9 to modify genomes. It describes: 1) Designing single guide RNAs (sgRNAs) to target specific gene loci using online tools; 2) Constructing plasmids expressing Cas9 and sgRNAs; 3) Validating plasmid function using assays like Surveyor nuclease; and 4) Transfecting cells, isolating clones, and further validating genome edits through sequencing. The goal is to use this method to precisely modify genomes for research applications.
Defined, consistent quality: The only all-in-one solution to simplify algae engineering:
GeneArt® Algae Engineering Kits for rapid production. Previously, algae research and production labs relied on poorly characterized, non-optimized cell stocks and cloning tools for their work. Preparing growth medium was convoluted and time-consuming, and growth rates and yields from the transformed cells were disappointing. New GeneArt® Algae Engineering Kits for Chlamydomonas reinhardtii and Synechococcus elongatus are the first commercially available genetic modification and expression systems for photosynthetic microalgae. These kits are designed for rapid scale-up and production and consistent, defined quality.
Next-generation genomics: an integrative approachHong ChangBum
This document summarizes a presentation on next-generation genomics and integrative analysis. It discusses the types of genomic data available from techniques like genome sequencing, RNA sequencing, ChIP-seq, and epigenomics. It explains that integrative analysis can help annotate functional features, infer variant function, and understand gene regulation. Approaches to integration include data reduction, unsupervised clustering, and supervised Bayesian networks. Large-scale datasets can be accessed through browsers, add-ons, and standalone tools to generate novel hypotheses. Future work includes more integrated community resources with search capabilities.
Lessons learned from high throughput CRISPR targeting in human cell linesChris Thorne
In just a short period of time CRISPR-Cas9 technology has revolutionized the field of genome editing, and taken the scientific community by storm. Already our understanding of how best to apply this technology has advanced significantly and almost every week new publications appear showcasing its application in basic and translational research.
While CRISPR-Cas9 is applicable across many different cell types, we have found it particularly suited for genome editing in near-haploid human cell lines. This has allowed us to establish a robust pipeline for the inactivation of non-essential genes at unprecedented scale and efficiency.
We have now knocked out over 1500 human genes and have generated a resource that is, to the best of our knowledge, the largest collection of human knockout cell lines available, covering comprehensive subsets of genes clustered by biological pathway (e.g. the autophagy pathway, the JAK/STAT pathway) or by phylogenetic relationship (e.g. kinases, bromodomain-containing proteins).
In this talk we will discuss how, through more than 1500 genome editing experiments, we have started to unravel some of the general principles governing the use of CRISPR-Cas9 in mammalian cells. For example, we have analyzed the impact of variation in the guide RNA sequence on Cas9 cleavage efficiency and characterized the mutational signature arising from CRISPR-Cas9 cleavage.
We will also highlight (with examples) how these learnings are now being applied to introduce other genomic modifications in a high throughput manner, including chromosomal deletions, translocations, point mutations and endogenous gene tags.
DNA Analysis - Basic Research : A Case StudyQIAGEN
The document discusses using the QIAxcel system for analyzing DNA fragments from genetic analyses like genotyping and mapping mutant genes. It provides examples of using the QIAxcel to genotype Arabidopsis thaliana plants with CAPS markers for a pks3 mutant allele. The system allows accurate sizing of DNA fragments in a high-throughput manner to unambiguously identify genotypes. It also discusses using the QIAxcel to analyze alternative splicing patterns regulated by the Tra2β protein to identify roles of its domains. The system provides rapid, sensitive and reproducible analysis of splicing patterns.
Next generation sequencing & microarray-- Genotypic TechnologyGenotypic Technology
Greetings from Genotypic Technology, Bangalore (www.genotypic.co.in). We are a 13 year old genomics and bioinformatics company ( 65+ employees, Service. Products and R & D) based in Bangalore, India, primarily working on applications of Microarrays and Next Generation Sequencing in Human Health and Disease, including in Molecular Diagnostics, Prognosis, Therapeutics, Vaccine Research, Basic Science Research, Veterinary Science, Agriculture, Industrial Biotechnology, Microbial Genetics and more.
Our major strength is in customized genomics solutions, particularly in your field, we can develop panel of markers for specific diseases, optimize, validate and help commercialize on open platforms or specific instrument platforms- in microarrays and sequencing based methods/ assays. We can also use genomic markers to aid in treatment of specific disease using personalized medicine approaches. All this can be done on a comprehensive end-to-end manner in our company as we have a very good blend of molecular biology and bioinformatics with totally 6 Ph.Ds. We work closely with Agilent's R &D as their partner.
PerkinElmer provides end-to-end next generation sequencing (NGS) services from sample intake to data analysis. Their CLIA-certified sequencing laboratory is staffed by expert scientists with decades of experience in genomics who deliver consistently high quality sequencing results. PerkinElmer offers sequencing, library preparation, capture, bioinformatics analysis, and professional consulting services to build customized NGS solutions that meet customers' specific needs and requirements.
The CRISPR/Cas9 system has emerged as one of the leading tools for modifying genomes of organisms ranging from E. coli to humans. Additionally, the simple gene targeting mechanism of CRISPR technology has been modified and adapted to other applications that include gene regulation, detection of intercellular trafficking, and pathogen detection. With a wealth of methods for introducing Cas9 and gRNAs into cells, it can be challenging to decide where to start. In this presentation, Dr Adam Clore describes the CRISPR mechanism and some of the most prominent uses for CRISPR, along with methods where IDT technologies can assist scientists in designing, testing, and executing a variety of CRISPR-mediated experiments. For more informaton, visit: http://www.idtdna.com/crispr
IDT provides oligonucleotides and panels for targeted sequencing including stocked and custom gene panels. Their panels include 264 genes for acute myeloid leukemia, 127 genes for pan-cancer analysis, and 4503 genes for inherited diseases. IDT probes are individually synthesized and quality controlled before being pooled. Universal blockers improve on-target rates by blocking adapter participation. Additional services include custom barcoded adapters and gBlocks fragments for quality control.
Targeted Breeding Applications of CRISPR-CasKate Barlow
Doane Chilcoat, Director, Applied Technology Systems, DuPont Pioneer
CRISPR-Cas as an advanced plant breeding tool is a more efficient way to improve plants and help farmers produce more and better food, with fewer resources. The superior properties of CRISPR-Cas allows DuPont Pioneer scientists to develop innovative and sustainable seed products for growers similar to those realized through conventional plant breeding, but with even greater efficiency, accuracy and quality. Pioneer is leading the application of this tool to develop customized agriculture solutions. In this talk, potential product targets of this promising technology will be discussed. Approaches to fostering social license and developing an open innovation model for CRISPR-Cas will also be reviewed.
Genome engineering using CRISPR/Cas9 has several advantages over traditional gene targeting methods: it is faster, more precise, applicable to many species, and less expensive. CRISPR/Cas9 uses the Cas9 nuclease guided by a single guide RNA to introduce double-strand breaks at targeted genomic loci. This can generate gene knockouts through error-prone non-homologous end joining or allow for targeted insertions and modifications through homology-directed repair. While CRISPR/Cas9 has great potential, careful design of guide RNAs and donor templates is needed to minimize off-target effects.
Recent breakthroughs in genome editing technology have led to a rapid adoption that parallels that seen with RNAi. And like RNAi, these methods are taking the scientific world by storm, with high profile publications in fields as diverse as HIV treatment, stem cell therapy, food crop modification and drug development to name but a few.
Critically, the endogenous modification of genes enables the study of their function in a physiological context. It also overcomes some of the artefacts that can result from established techniques such as transgenesis and RNAi, which have mislead researchers with false positives or negatives. Until recently however genome editing required considerable technical expertise, and consequently was a relatively niche pursuit.
In this talk we will look at how the latest developments in genome editing tools have changed this, with improvements in both ease-of-use and targeting efficiency, as well as a concomitant reduction in costs opening up these approaches to the wider scientific community.
Rapid adoption of the CRISPR/Cas9 system has for example led to a long list of organisms and tissues in which genetic changes have been made with high efficiency. Other technologies such as recombinant adeno-associated virus (rAAV) offer further precision, stimulating the cell’s high-fidelity DNA repair pathways to insert exogenous sequence with unrivalled specificity. Targeting efficiency can be improved still further by using the technologies in combination – genome cutting induced by CRISPR can significantly enhance homologous recombination mediated by rAAV.
Despite these rapid advances, some pitfalls remain, and so we’ll discuss some of the key considerations for avoiding these, ranging from simply picking the right tool for the job to designing an experiment that maximises chances of success.
Finally we’ll look at how genome editing is being applied to both basic and translational research, and in both a gene-specific and genome wide manner. For the study of disease associated genes and mutations scientists can now complement wide panels of tumour cells with genetically defined isogenic cell pairs identical in all but precise modifications in their gene of interest. The ease-of-design and efficiency of the CRISPR system is also being exploited for genome wide synthetic lethality screens, facilitating rapid drug target identification with significantly reduced risk of false negatives and off-target false positives. And again, further synergies are achieved when these approaches are combined to look for potential synthetic lethal targets in specific genomic contexts.
The document summarizes molecular characterization of Puccinia striiformis f.sp. tritici (Pst) isolates from Western Canada. Pst isolates were sequenced using Illumina platforms and assembled de novo. Phylogenetic trees were constructed based on rRNA sequences and whole genome assemblies. Comparisons between old isolates from 1990-1993 and new isolates from 2007-2012 identified unique and enriched gene sequences, suggesting genome reorganization in Pst. Functional annotation revealed differences in biological processes between old and new isolates, such as transport and response to exogenous molecules in new isolates.
Fast and Efficient Post-Bisulfite-Seq Library Construction with QIAseq Ultral...QIAGEN
The document describes a new method called post-bisulfite next-generation sequencing library construction (PBLC) for whole genome bisulfite sequencing (WGBS) that overcomes challenges with traditional WGBS workflows. PBLC involves bisulfite treatment before library preparation, which fragments the DNA so no additional fragmentation is needed. This reduces workflow time and enables epigenomic studies from lower DNA inputs. Comparison studies show PBLC produces higher library yields than traditional WGBS and comparable sequencing results, making it a more efficient and sensitive method for WGBS.
This document provides an overview of CRISPR/Cas9 genome editing. It discusses the history and limitations of prior genome engineering techniques like recombinant DNA and zinc finger nucleases. It then explains how CRISPR/Cas9 works as a RNA-guided DNA endonuclease and how this allows it to efficiently and specifically edit genomes. The document outlines several applications of CRISPR/Cas9 like generating knockout animals and cell lines. It also notes some concerns about using the technique for human genome editing.
This document discusses genome editing using the CRISPR-Cas9 system. It begins by introducing three main genome editing technologies - zinc-finger nucleases, TALENs, and the CRISPR-Cas9 system. It then describes the key events in the discovery of CRISPR-Cas9, including its origins as a bacterial defense system. The document outlines the main components of the CRISPR-Cas9 system, including crRNA, tracrRNA, sgRNA, and Cas9. It also summarizes the two main steps in genome editing using CRISPR-Cas9 - knocking out genes and DNA repair. The document concludes by discussing opportunities for applying CRISPR-Cas9 technology across various
The CRISPR/Cas9 system has emerged as one of the leading tools for modifying the genomes of organisms ranging from E. coli to humans. In this presentation, we discuss various methods for generating the crRNA and tracrRNA components that are required for guiding the Cas9 endonuclease to genomic targets. You will also learn how to optimize a new 2-part CRISPR RNA system from IDT that offers multiple benefits over other technologies.
Multi Target Gene Editing using CRISPR Technology for Crop ImprovementTushar Gajare
This document provides an overview of a presentation on using CRISPR technology for multi-target gene editing in crop improvement. It begins with an introduction to genome editing and CRISPR-Cas9. It then discusses the CRISPR system, how CRISPR-Cas9 works, its history and applications for crop improvement including case studies in maize with high mutant efficiencies and targeted mutagenesis of multiple genes. The presentation covers advantages and limitations of the technology as well as future prospects.
The document provides an overview of gene knockout and homology-directed repair using CRISPR. It discusses designing guide RNAs and comparing delivery methods like lipofection, electroporation, and microinjection. It also covers designing repair templates for homology-directed repair to insert or change DNA sequences. Optimization of guide RNAs, delivery method, and repair template design can improve genome editing efficiency.
This document summarizes QIAGEN's products for sample preparation, targeted sequencing, and single-cell analysis across various areas of biomedical research including liquid biopsy, circulating tumor cells, and gene expression profiling. Key products mentioned are the QIAseq cfDNA All-in-One Kits for streamlined library preparation from plasma/serum, QIAseq Targeted DNA/RNA Panels for digital sequencing of genomic regions or genes, and QIAseq FX Single Cell DNA/RNA Library Kits for cell-to-library workflows from isolated single cells.
Ernesto Picardi – Bioinformatica e genomica comparata: nuove strategie sperim...eventi-ITBbari
Bioinformatica e genomica comparata: nuove strategie sperimentali e computazionali per la produzione e analisi di dati NGS finalizzati a sviluppare processi e prodotti innovativi per la salute dell’uomo, l’ambiente e l’agroalimentare.
CRISPR-Cas9 is a powerful tool for genome engineering. The document provides guidance on using CRISPR-Cas9 to modify genomes. It describes: 1) Designing single guide RNAs (sgRNAs) to target specific gene loci using online tools; 2) Constructing plasmids expressing Cas9 and sgRNAs; 3) Validating plasmid function using assays like Surveyor nuclease; and 4) Transfecting cells, isolating clones, and further validating genome edits through sequencing. The goal is to use this method to precisely modify genomes for research applications.
Defined, consistent quality: The only all-in-one solution to simplify algae engineering:
GeneArt® Algae Engineering Kits for rapid production. Previously, algae research and production labs relied on poorly characterized, non-optimized cell stocks and cloning tools for their work. Preparing growth medium was convoluted and time-consuming, and growth rates and yields from the transformed cells were disappointing. New GeneArt® Algae Engineering Kits for Chlamydomonas reinhardtii and Synechococcus elongatus are the first commercially available genetic modification and expression systems for photosynthetic microalgae. These kits are designed for rapid scale-up and production and consistent, defined quality.
Next-generation genomics: an integrative approachHong ChangBum
This document summarizes a presentation on next-generation genomics and integrative analysis. It discusses the types of genomic data available from techniques like genome sequencing, RNA sequencing, ChIP-seq, and epigenomics. It explains that integrative analysis can help annotate functional features, infer variant function, and understand gene regulation. Approaches to integration include data reduction, unsupervised clustering, and supervised Bayesian networks. Large-scale datasets can be accessed through browsers, add-ons, and standalone tools to generate novel hypotheses. Future work includes more integrated community resources with search capabilities.
Lessons learned from high throughput CRISPR targeting in human cell linesChris Thorne
In just a short period of time CRISPR-Cas9 technology has revolutionized the field of genome editing, and taken the scientific community by storm. Already our understanding of how best to apply this technology has advanced significantly and almost every week new publications appear showcasing its application in basic and translational research.
While CRISPR-Cas9 is applicable across many different cell types, we have found it particularly suited for genome editing in near-haploid human cell lines. This has allowed us to establish a robust pipeline for the inactivation of non-essential genes at unprecedented scale and efficiency.
We have now knocked out over 1500 human genes and have generated a resource that is, to the best of our knowledge, the largest collection of human knockout cell lines available, covering comprehensive subsets of genes clustered by biological pathway (e.g. the autophagy pathway, the JAK/STAT pathway) or by phylogenetic relationship (e.g. kinases, bromodomain-containing proteins).
In this talk we will discuss how, through more than 1500 genome editing experiments, we have started to unravel some of the general principles governing the use of CRISPR-Cas9 in mammalian cells. For example, we have analyzed the impact of variation in the guide RNA sequence on Cas9 cleavage efficiency and characterized the mutational signature arising from CRISPR-Cas9 cleavage.
We will also highlight (with examples) how these learnings are now being applied to introduce other genomic modifications in a high throughput manner, including chromosomal deletions, translocations, point mutations and endogenous gene tags.
DNA Analysis - Basic Research : A Case StudyQIAGEN
The document discusses using the QIAxcel system for analyzing DNA fragments from genetic analyses like genotyping and mapping mutant genes. It provides examples of using the QIAxcel to genotype Arabidopsis thaliana plants with CAPS markers for a pks3 mutant allele. The system allows accurate sizing of DNA fragments in a high-throughput manner to unambiguously identify genotypes. It also discusses using the QIAxcel to analyze alternative splicing patterns regulated by the Tra2β protein to identify roles of its domains. The system provides rapid, sensitive and reproducible analysis of splicing patterns.
Next generation sequencing & microarray-- Genotypic TechnologyGenotypic Technology
Greetings from Genotypic Technology, Bangalore (www.genotypic.co.in). We are a 13 year old genomics and bioinformatics company ( 65+ employees, Service. Products and R & D) based in Bangalore, India, primarily working on applications of Microarrays and Next Generation Sequencing in Human Health and Disease, including in Molecular Diagnostics, Prognosis, Therapeutics, Vaccine Research, Basic Science Research, Veterinary Science, Agriculture, Industrial Biotechnology, Microbial Genetics and more.
Our major strength is in customized genomics solutions, particularly in your field, we can develop panel of markers for specific diseases, optimize, validate and help commercialize on open platforms or specific instrument platforms- in microarrays and sequencing based methods/ assays. We can also use genomic markers to aid in treatment of specific disease using personalized medicine approaches. All this can be done on a comprehensive end-to-end manner in our company as we have a very good blend of molecular biology and bioinformatics with totally 6 Ph.Ds. We work closely with Agilent's R &D as their partner.
PerkinElmer provides end-to-end next generation sequencing (NGS) services from sample intake to data analysis. Their CLIA-certified sequencing laboratory is staffed by expert scientists with decades of experience in genomics who deliver consistently high quality sequencing results. PerkinElmer offers sequencing, library preparation, capture, bioinformatics analysis, and professional consulting services to build customized NGS solutions that meet customers' specific needs and requirements.
Exploring new frontiers with next-generation sequencingQIAGEN
The document describes several next-generation sequencing products from QIAGEN, including the QIAseq Targeted RNA Panels, QIAseq Targeted RNAscan Panels, QIAseq Targeted DNA Panels, QIAseq 1-Step Amplicon Library Kit, QIAseq Ultralow Input Library Kit, and QIAseq cfDNA All-in-One Kit. These products are designed to simplify NGS workflows and provide high-quality libraries from low input samples in order to maximize insights from applications such as single-cell analysis, liquid biopsies, and metagenomics.
The QIAGEN Service Core provides a full service for genomic and expression analysis using cutting-edge tools. It accepts various sample types and performs experiments, such as pathway-focused analysis, whole genome analysis, and real-time PCR, using reagents and equipment. This allows customers to save costs by avoiding investing in their own resources and saves time by outsourcing experimental work. A variety of nucleic acid and data analysis services are available.
GenScript is a world-leading biology CRO with over 1,000 employees and facilities in New Jersey, Nanjing, Paris, and Tokyo. It provides a range of custom biology services including gene synthesis, protein expression, antibody production, and assay development to support drug discovery. GenScript has significant production capacity and aims to remove bottlenecks in early-stage drug development through its integrated service offerings.
GenScript is a world-leading biology CRO with over 1,000 employees and facilities in New Jersey, Nanjing, Paris, and Tokyo. It provides custom gene synthesis, protein and peptide services, antibodies, and assay development. GenScript has a drug discovery platform and removes bottlenecks in early drug discovery through custom bio-reagent preparation and screening.
GenScript is a biotechnology company founded in 2002 that provides gene synthesis, protein and antibody services. It has over 800 employees across facilities in the US, China, France and Japan. GenScript is the largest provider of custom gene synthesis in the US and offers services including gene synthesis, cloning, mutagenesis, and plasmid preparation. It also provides protein and antibody services such as expression, purification, production and development of monoclonal and polyclonal antibodies. GenScript focuses on intellectual property protection and being a reliable partner for its clients in the pharmaceutical and biotech industries.
GenScript is a biotechnology company founded in 2002 that provides gene synthesis, protein and antibody services. It has over 800 employees across facilities in the US, China, France and Japan. GenScript is the largest provider of custom gene synthesis in the US and offers services including gene synthesis, cloning, mutagenesis, and plasmid preparation. It also provides protein and antibody services such as expression, purification, production and development of monoclonal and polyclonal antibodies. GenScript focuses on intellectual property protection and being a reliable partner for its pharmaceutical and biotech clients.
Dr. Chris Lowe presented on Horizon Discovery's precision genome editing platform called GENESISTM. The presentation discussed optimizing GENESISTM by combining CRISPR and rAAV technologies to improve gene targeting efficiency. Custom cell line development services are offered to modify genes of interest in various cell lines for applications such as generating disease models and studying drug sensitivity. Key considerations for successful gene editing experiments include factors like gene/cell line selection, gRNA design/activity, donor design, screening/validation approaches. Case studies demonstrated applications of engineered cell lines.
Casey Molecular Diagnostic Laboratory has developed a new next-generation sequencing panel method using simplex PCR reactions run on a chip with over 5,000 individual wells, with each primer set duplicated to avoid allele dropout. This new approach offers unmatched target enrichment of over 98% and average coverage over 500x. Every novel variation is manually checked to avoid false positives or negatives. While other labs sequence more broadly, Casey believes their targeted approach focusing on relevant genes optimizes mutation detection.
The document summarizes a presentation on bioinformatics case studies focusing on epigenetics and personal genomics. It discusses DNA methylation and its role in cancer development. It also describes how next-generation sequencing can be used to identify epigenetic biomarkers for clinical use. Finally, it addresses issues around personal and recreational genomics, including increasing access, educating users, and protecting individual privacy and rights.
Sirion offers genetically modified cell services including cell line optimization, immortalization, and gene overexpression or silencing using viral vectors like adenovirus and lentivirus. They guarantee delivery of any gene to any mammalian cell type and greater than 90% knockdown maintained over experimental timeframes. Sirion claims to provide customized genetically modified cells, viruses, and services in weeks rather than the months needed for in-house solutions.
Advanced Genome Engineering Services and Transgenic Model Generation
at MSU’s Transgenic and Genome Editing Facility
Huirong Xie, Elena Demireva, Nate Kauffman, Richard Neubig
ArrayGen is a global bioinformatics company that provides one-stop solutions for NGS data analysis, microarray data analysis, and bioinformatics application development. It offers services such as custom array design, NGS data validation, gene expression analysis, and bioinformatics services. ArrayGen also develops products like Agilent catalog microarrays and the ANG genome browser to enable genomics research.
ArrayGen is a global genomics service provider company which is a one stop solution for Genomics Bioinformatics algorithm development and data analysis. ArrayGen's focus on Genomics, NGS data analysis, custom array design, microarray data analysis thus cover a vast majority of Bioinformatics applications in the life sciences sector.
In this slides the topic that which is discussed is "How PCR is involved in identification of Genotype"
I hope this will Help you in your presentation work.
"PCR can be used in identification of genotype."
mRNA (messenger RNA), a single-stranded molecule, is the genetic coding templates used by the translational machinery. mRNA leaves the cell nucleus and moves to the cytoplasm where the translation machinery makes proteins bind to these mRNA molecules and read the code on the mRNA to make a specific protein.https://mrna.creative-biolabs.com/mrna-services.htm
This document discusses oncogenomics and cancer genomics technologies. It provides an overview of oncogenomics, the types of DNA biomarkers studied including mutations, and experimental strategies used for cancer genomics research. Key techniques discussed are next-generation sequencing, quantitative PCR (qPCR), and mass spectrometry. The document compares different technologies for mutation detection and profiling and their sensitivities. It also outlines the specifications and pipeline for developing a qPCR-based somatic mutation assay.
1. FindYour Place in Genome Engineering
Transposagen Biopharmaceuticals, Inc. 1-844-GEN-EDIT
info@transposagenbio.com
www.transposagenbio.com
Technologies,Tools & Services Brochure
piggyBac™ • XTN™ TALENs • NextGEN™ CRISPR • Footprint-Free™ Gene Editing
• Custom Vectors • Cell Line Engineering • Custom Animal Models and More!
Connect with us for the latest updates on gene editing
technologies, products, services and promotions!
2. Transposagen Biopharmaceuticals, Inc. www.transposagenbio.com
About Us
Welcome to Transposagen Biopharmaceuticals, Inc., a worldwide leader in genome engineering technologies and
services with applications in therapeutics, research & drug discovery, bioproduction, clinical genetic testing and
agriculture. Our products and services include Footprint-Free™ Gene Editing, NextGEN™ CRISPR, XTN™ TALENs,
and custom cell lines, stem cells, and animal models. Our unique genome engineering capabilities allow for the
creation of nearly any genetic modification in any genome.
• Anyone can cut DNA (even site-
specifically)
• Our technology is the cleanest
gene editing technology in the
world
• We are the only company in the
world that can edit as little as a
single nucleotide without any
unwanted changes and with the
ability to select for rare events
• We have a 20-year patent
position protecting this
technology
Custom reagents and
technology portfolio
Cell and
animal model
engineering
services
Drug development
partnerships
What Differentiates Us WhatWe Offer
• Drug Discovery & Development
Elucidating gene function & regulation
Target discovery
Disease models
Toxicology & reporter lines
Isogenic control lines & diagnostics
• Bioproduction
Host engineering for improved
production and function
Stable cell lines and pools
• Therapeutics
Gene therapy
Cell therapy – ex vivo cell therapy
Why Genome Engineering
1-844-GEN-EDIT
info@transposagenbio.com
3. The piggyBac™DNA Modification System
Transposagen Biopharmaceuticals, Inc. www.transposagenbio.com
Transposagen is proud to provide the industry’s most efficient non-viral stable transfection & transgenesis
technology. Choose any of our off-the-shelf transposons and stable transfection kits or custom transposon services
in order to facilitate stable transfection and transgenesis. Simple, one step protocol from transfection to stable cell
lines.
How ItWorks
Step 1. Clone your custom cargo into
piggyBac™ transposon vector.
Step 2. Transfect the piggyBac transposon
containing inserted cargo along with the
piggyBac™ transposase (plasmid DNA or
mRNA) into target cell.
Step 3. piggyBac™ transposase recognizes
PB transposon sequences (ITR’s), cuts
transposon, carrying the cargo out of the
plasmid and integrates it stably in the
genome.
Advantages
No size limit- up to 250 kb has been published
80-100% stable integration efficiency
Effective on all mammalian genome
Precise Excision
Non- Toxic and non-mutagenic
Applications
• Stable cell line creation
• Stem cell research: reprogramming, differentiation
and selection.
• Protein production
• Cell and gene therapy
• Drug Screens
• RNAi: inducible and reversible knockdown in cells and
animals models
Using piggyBac™ for Stable Transfection & Transgenesis
PB- GOLD™ Stable Transfection Kit Contents
• hyperactive super piggyBac™
transposase (sPBo) plasmid
• lipofection reagents
• positive control plasmid
Unmatched Stable Expression
THP-1 Reporter Lines
Red = negative control
Green = initial transfection expression
Blue = expression after 10 months
Mossine et al. 2013
Outperforms Viral & Plasmid Methods
1-844-GEN-EDIT
info@transposagenbio.com
4. Transposagen Biopharmaceuticals, Inc. www.transposagenbio.com
piggyBac™ Excision-only Transposase
piggyBac™ excision only transposase allows you to
remove cargo from the in a Footprint-Free™, seamless or
scarless manner. Use for:
Footprint-Free Gene Editing
Phenotype Reversion Studies
Custom Services
Transposagen provides everything from vector to full cell
line or animal model creation.
• Custom Transposon vectors
• Custom Cell Lines
• Custom Transgenic animal models
piggyBac™ Footprint-Free™ Excision
PB-x GOLD™ Excision-only Transfection Kit
Contents
• Excision-only piggyBac™ transposase
(PBx) plasmid
• lipofection reagents
• positive control plasmid
RapidTransgenesis
Pettitt et al 2013
Genetic Screen’s & Phenotype Reversion Studies
Demonstration of the piggyBac™
capability to produce large stable
mutant libraries in forward genetic
gene trapping screens. Genes
involved in compound (Olaparib)
toxicity resistance were discovered
(left). Followed by phenotype
reversion back to sensitivity with
excision only piggyBac™ (right)
Green = wild type sensitive cells
Blue = resistant cells with PB gene trap insertions
Grey = reverted excised clones
Blue= resistant cells with gene trap insertion
Green = wild type sensitive cells
Red = negative control
Off-the-shelf vectors
Transposon backbone vectors for cloning your cargo of
interest. The piggyBac™ ITRs flanking core insulators and a
multiple cloning site (MCS). Our backbones come with a
variety of promoters, markers and other features.
1-844-GEN-EDIT
info@transposagenbio.com
5. Site-Specific Nucleases for Gene Editing
Transposagen Biopharmaceuticals, Inc. www.transposagenbio.com
Site-specific nucleases enable researchers to accurately cut any DNA sequence, allowing for the creation of
gene knock-outs, knock-ins and single base pair edits. Transposagen offers the widest variety of nucleases
including our XTN™ TALENs and the latest dimeric RNA-guided NextGEN™ CRISPR. Not only can Transposagen
provide you with the latest DNA editing technologies, but we have the fastest delivery time in the industry.
Genome Modification Applications
Avoid Off-Targets With NextGEN™ CRISPR
Some nucleases, especially the original CRISPR/Cas9
technology have the unfortunate consequence of off-
target mutations. The NextGEN™ CRISPR technology is the
first truly dimeric RNA-guided FokI nuclease which
diminishes these concerns. The NextGEN™ CRISPR system
empowers researchers to engineer the genome with the
highest fidelity without compromising the budget.
Affordable
Fast
High efficiency
Able to multiplex
Highest fidelity; no detectable off-
targets by deep sequencing
methods
• Gene knock-out and knock-ins
• Single base-pair gene editing
• Mutation correction or creation
• Cell line & animal model engineering
• Validation of GWAS studies and patient
clinical trial selection
• Cellular therapies
NextGEN™ architecture. The NextGEN™ CRISPR utilizes the dimerization requirement of FokI for improved specificity.
By introducing site-specific double-strand breaks,
nucleases promote the NHEJ and HR pathways which
enable gene editing.
Custom Reagents
1-844-GEN-EDIT
info@transposagenbio.com
6. Succeed With XTN™ TALENs
Transposagen Biopharmaceuticals, Inc. www.transposagenbio.com
Transposagen is an experienced XTN™ TALEN producer. In fact, we use XTN™ TALENs for our own in-house
projects for producing genetically engineered rats and cells lines. Our XTN™ TALENs have to be of the highest
quality and activity in order to achieve results for our customers. We deliver our commercial XTN™ TALENs with
the same standards in order to help you achieve your gene editing goals.
Get the best – Not all TALENs are equal.
Transposagen uses the superior protein
engineering to deliver high performance TALENs.
Enhanced targeting, better promoters, variable
repeats and codon optimization make all the
difference.
Have them fast – We utilize our own XTN™ high-
throughput production system to deliver your
TALENs quickly.
Target your gene more often – XTN™ TALENs have
a wide targeting range in the genome making all
types of research possible.
Acquire results – Higher efficiencies translate to
better results.
Stay out of trouble – XTN™ TALENS have minimal
off-target effects and low toxicity.
Site-Specific Nuclease Product & Service Options
Transposagen has a number of innovative site-specific nuclease service programs designed to fit the needs and budget
of any research or drug discovery project. We offer everything from off-the-shelf gene editing tools to the most
complex animal model creation services.
Advantages of XTN™TALENs
→ Value™ Program – Receive custom NextGEN™ CRISPR and/or XTN™ TALEN plasmids sequenced verified and
transfection ready.
→ Guaranteed Success™ Program – Two custom NextGEN™ CRISPR and/or XTN™ TALEN plasmids sequenced
verified and transfection ready. If the first pair of nucleases do not cut at your target you build more for FREE.
→ Gene Editing Kit – Two custom NextGEN™ CRISPR and/or XTN™ TALEN plasmids and custom donor vector
sequenced verified and transfection ready.
→ mRNA Production Service – High quality mRNA production for any custom NextGEN™ CRISPR and/or XTN™
TALEN.
TakeYour Nucleases ToThe Next Level
Ask your local Gene Editing Specialist about our unique Footprint-Free™ Gene Editing piggyBac™ reagents and services.
Combine NextGEN™ CRISPR With XTN™ TALENs
Unlike other providers in the industry, Transposagen offers both CRISPR and TALEN technologies to researchers. Since
each nuclease has inherent sequence requirements, the option of both provides a wider targeting range than alone.
Combine both nucleases in a single transfection for even higher chances of successful gene targeting!
Custom Reagents
XTN™ TALEN Rat Rosa26 targeting compared
to a competitor TALEN producer.
XTN™ Competitor
Bands indicate
activity by Cel1
assay
Evidence of XTN™ Efficiencies
• Hundreds of successful projects in cell line and animal model
creation.
• Direct comparisons between our XTN™ TALENs and competitor
producers.
1-844-GEN-EDIT
info@transposagenbio.com
7. Footprint-Free™ Gene Editing
Transposagen Biopharmaceuticals, Inc. www.transposagenbio.com
Transposagen’s Footprint-Free™ gene editing kits & services combine the industry’s most precise, efficient, and
flexible site-specific nucleases, NextGEN™ CRISPRs and XTN™ TALENs, with our exclusive piggyBac™ transposon
system - the only commercially available method capable of seamless excision of resistance or reporter genes.
Footprint-Free™ Gene Editing allows users to increase efficiencies by utilizing selection enrichment for gene edits,
followed by “footprint-free” or “scarless” removal of the selection cassette, resulting in clean genome editing.
How ItWorks
1- Start with the most efficient &
precise targeted nucleases,
NextGEN™ CRISPR and/or XTN™
TALENs to stimulate homologous
recombination at the target site.
2- Use a Footprint-Free™ Gene
Editing donor plasmid from our
Multivector™ portfolio for
targeted integration and selection
enrichment of the desired gene
edit.
3- Following selection Excision
Only piggyBac™ (PBx) removes
markers, leaving behind a scarless
correctly edited genomic DNA.
Genome Modification Applications
• Gene editing down to a single base-pair –
mutation correction or creation in cell lines with
key advantages in difficult to transfect cells such
as stem cells (iPS and ES cells) and primary cells.
• Targeted knock-ins, reporter lines and
humanizations.
• Targeted knockouts and knockout reversions –
introduce a disruption (selection marker) in the
coding region, following phenotypic analysis
remove the disruption Footprint-Free™ for
phenotype reversion studies.
Product & Service Options
Transposagen provides a range of services from the off-
the-shelf Multivector™ portfolio to custom gene editing
kits. Footprint-Free™ cell lines and animal model
services are also available.
Multivector™ Catalog #
PB-MV1Puro-TK
PB-MV1Neo-TK
PB-MV1CMV-GFP-Puro
PB-MV1Neo
PB-MV1Hygro-TK
Technology Applications
1-844-GEN-EDIT
info@transposagenbio.com
8. Project Applications
Transposagen Biopharmaceuticals, Inc. www.transposagenbio.com
Cell line engineering - Access to cell lines and stem cells from genetic disease
or modified backgrounds is becoming more common and important for in vitro
(cell based) modeling. Genomes vary in population, including SNPs in coding
regions and regulatory elements as well as copy number. When compared to
“control or regular stem cells” these natural variations result in
misinterpretation of in vitro phenotypes, making creation of isogenic lines used
as a reference for genetically modified or disease lines necessary for quality
results.
Advantages in Stem Cells - Genome engineering in common transfection cell
lines such as HEK293 occurs at high efficiencies. However, stem cells are less
amendable to genome modification due to reduced transfection efficiencies
and potentially different DNA repair responses. Therefore, selection for
genome modifications through Footprint-Free™ Gene Editing is a key benefit
for stem cell engineering.
Cellular therapies – treatments for a broad range of disorders with clinical
capabilities (no off-target mutations). piggyBac™ and Footprint-Free™ Gene
Editing have been used in CAR T-cell therapy studies as well as correction of
multiple disease mutations.
Isogenic cell line
creation
GWAS Study
Validation
Target discovery &
validation
Patient clinical trial
selection
Cell & GeneTherpay
Animal model
creation
Selection vs. single-stranded DNA oligonucleotide
(ssODN) mediated gene editing: One alternative to
selection is the use of ssODNs which are typically 60-120
bases with the donor substitution (base-pair edits)
flanked by homologous sequences on each side.
Inefficiencies require a large number of clones to be
screened resulting in project timeline delays and cost
increases. Footprint-Free™ selection enrichment allows
simpler easier screening, for faster and more cost
effective genome editing.
Advantages of Footprint-Free™ Gene Editing
Why selection? Site-specific nucleases increase the efficiency of targeted knock-ins or gene edits significantly, but
in many cases efficiencies may not be high enough for clone isolation. Therefore, the ability to select for desired
mutations and rare events is important for efficient gene editing.
Footprint-Free™ Gene Editing vs. CRE/loxP
recombinase: Recombinase methods remove drug
selection markers from the genome by catalyzing
recombination between target sites (known as loxP
sites). However, this process is not clean and leaves
“footprints” or “scars” in the genome which have
been shown to disturb splicing elements or
alter/silence gene expression. In addition,
recombinases recognize “cryptic” loxP sites and
produce off-target mutations, compromising results.
Footprint-Free™ gene editing does not have these
adverse effects providing precise and efficient
genome editing for quality results.
Access to Footprint-Free™
Gene Editing
1- Multivector™ Portfolio – generate
your own Footprint-Free™ gene editing
vectors
2- Custom Footprint-Free™ Gene
Editing Kits – Let us design and
produce the vectors for you
3- Custom Cell Line Engineering &
Animal Model Creation – Utilize
our expertise and outsource the
whole project
Technology Applications
1-844-GEN-EDIT
info@transposagenbio.com
9. Custom Cell Line Engineering
Transposagen Biopharmaceuticals, Inc. www.transposagenbio.com
Transposagen utilizes expertise and a diverse portfolio of gene editing tools including XTN™ TALENs, NextGEN™
CRISPRs, piggyBac™ transposons and Footprint-Free™ Gene Editing to deliver high quality custom engineered cell lines
Available Services
Stable cell lines (transgenic,
overexpression, inducible)
Knockout
Knock-in (humanization &
conditional)
Footprint-Free™ Gene Editing
Cell line characterization services
Custom Services
An example of our 4-step milestone pricing structure for
high efficiency knockout cell lines
Best Reagents
Your custom engineered cell line begins with the latest gene editing
reagents available. Transposagen uses TALEN and CRISPR
technologies, including the latest dimeric NextGEN™ CRISPR which has
been shown to have minimal off-target effects. Combined with
piggyBac Footprint-Free Gene Editing, Transposagen achieves the
highest efficiency.
Expertise
Transposagen has over 10 years of experience engineering custom cell
lines and animal models. We've successfully delivered projects to the
top 20 pharmaceutical companies and all the major academic
institutes. We have experience with many cell lines including iPS, SSCs,
CHO, HEK293, HeLa, ES, SH-SY5Y, HepG2, A549, MCF7, CACO-2,
RAW264.7, THP-1, Jurkat... to name a few!
Time
Generating a cell line can be time-consuming
and complex. By utilizing the piggyBac™ system,
Transposagen can delivery engineered cell lines
faster than anyone in the industry for even the
most difficult lines.
Accurate Pricing & Flexible Projects
No surprises. Prior to the start of any project,
Transposagen can accurately price out the cost
of your engineered cell line so you know what to
expect. All cell line projects are divided into
milestone payments which allows researchers to
start the service without paying for all of it
upfront. You only pay for the work we complete!
KeepYour Investment
The reagents we use to engineer your cell line
are yours to keep. Use them again in a different
cell line or take them to our animal model
services to maximize your research efforts.A demonstration of knockout
cell line creation at
Transposagen. Inactivating
the fucosylation system in
CHO cells for producing
afucosylated human
therapeutics. Antibodies
expressed from these host
cells were afucosylated and
demonstrated the expected
increased ADCC activity.
In collaboration with Aragen
Bioscience.
Targeting Fut8, the alpha1,6-fucosyltransferase gene
with XTN™TALENs in CHO Cells
1-844-GEN-EDIT
info@transposagenbio.com
10. Custom Animal Models
Transposagen Biopharmaceuticals, Inc. www.transposagenbio.com
Transposagen has the industry’s most versatile collection of genome engineering technologies as well as
expertise in rodent pronuclear injection (PNI) and stem cell engineering. These capabilities result in the most
flexible and robust custom model generation services in the industry. The more targeted and complicated your
therapeutics become, the more you need the right partner to advance your research. Working intimately with
our scientific team will help you bring innovative solutions and products to market faster.
Custom Services
Available Rat & Mouse Services
Knockout
Transgenic Overexpression
Conditional
Knock-in
Transgenic knockdown
Humanized
Disease Models
1-844-GEN-EDIT
info@transposagenbio.com
Genome Engineering Vector Design & Validation
+
PNI, ES or SSCs
High-speed, high-efficiency genome engineering
Rodent Model Creation Strategies
Method Species Targeted Knock-in
Limitation
Estimated Timeline
Pronuclear Injection (PNI) Mouse & Rat ~2kb 4 months
Embryonic Stem Cells (ES) Mouse None 8 months
Spermatogonial Stem
Cells (SSC)
Rat None 8 months
• Custom Engineered Rodents in as little as 4 months
• Spermatogonial Stem Cell (SSC) technology is the only commercial method for producing
large targeted knock-ins in rats
11. Transposagen Biopharmaceuticals, Inc. www.transposagenbio.com
FatRat™ Mc4r Knockout Obesity-Diabetes Model
Research Models
Featured Knockout Rat Models
Drug Metabolism
• Bcrp
Immune System
• Rag2 - SCID
Neuroscience –CNS
• SERT
• OPRL1
• Ghsr
• Nrg1
• Trpc4
• Pde4d
• Myo9a
Metabolic Disease/Obesity
• Mc4r – FatRat™
• Leptin
Cardiovascular
• Sod3
Cancer/Toxicology
• P53
1-844-GEN-EDIT
info@transposagenbio.com
Transposagen’s off the shelf TKO® and TGEM® rat models cover a range of disease and research applications. We
have partnered with world experts in over 40 collaborations worldwide in order to validate and characterize our
genetically engineered rat models. We have approximately 300 off-the-shelf models which is the world’s largest
commercially available collection. Our characterized genetically engineered models are broken down into the
research categories below.
• Most common monogenic obesity gene
• Obesity studies, including early and late onset obesity
• Diabetes research
• Metabolism studies
• Renal studies
• Altered Leptin Signaling Pathway
Rag2 –SCID Knockout Rat
• Cell sorting data demonstrates no mature T
and B cells
• Western blot /Immunochemistry indicates
no IgG
• Initial results indicate that the animals are
athymic
New SCID Rat for Human Xenograft
Tumor Models
Access to Research Models
Unlimited breeding licenses - breeding at the
customer’s in-house facility or a contract breeder of
choice. The license comes with at least 2-3
heterozygous breeding pairs.
Trial licenses- allow the customer to purchase a
limited breeding license to conduct pilot studies at
a lower purchase price.
Individual animals – may be available, please
inquire
12. Transposagen Biopharmaceuticals, Inc. 1-844-GEN-EDIT
info@transposagenbio.com
www.transposagenbio.com
Connect with us for the latest updates on gene editing
technologies, products, services and promotions!
Exon Exon Exon
One-Step Stable
Transfection & Transgenesis
Targeted Knockouts
Footprint-Free™
Gene Editing
NextGEN™ CRISPR
StableTransfection Kits &
piggyBac™ Transposon
XTN™TALEN
Knock-ins &Humanization
Gene Editing
Base-Pair Changes
Cell Line & Rodent Model Engineering Services
Your Next Discovery Awaits
Targeted Selection
Markers
With So Many Opportunities
What AreYou Waiting For?
Next-Generation Genome Engineering