SlideShare a Scribd company logo
1 of 5
CustomizedCustomizedCustomizedCustomized ServicesServicesServicesServices
ForForForFor StableStableStableStable Knock-DownKnock-DownKnock-DownKnock-Down orororor Over-ExpressionOver-ExpressionOver-ExpressionOver-Expression
CellCellCellCell LineLineLineLine GenerationGenerationGenerationGeneration
Stable cell lines with specific gene over-expression or knock-down are very
helpful in gene function analysis, target discovery, target validation, assay
development, and compound screening. However, generation of stable cell
lines is a time-consuming and expensive process. In the last 4 years,
Shanghai Genomics has established a team of experts for stable cell line
generation. We have successfully generated hundreds of stable cell lines to
meet the need of our customers.
MethodMethodMethodMethod
Shanghai Genomics typically uses non-viral (liposome) and viral (lentivirus)
platforms for stable cell line generation. For some cell lines, like HEK293
and HeLa, the liposome method can achieve high transfection efficiency.
Compared to liposomal or other chemical vehicles, a lentiviral approach has
three distinct advantages:
1. The pseudotype virus has a broad range of host cell lines and is able to
infect both dividing and non-dividing cells, making it especially suitable for
some differentiated cell lines such as neurons, macrophages, hematopoietic
stem cells, retinal photoreceptors and muscle and liver cells.
2. The virally encoded integrase greatly facilitates exogenous gene
integration that ensures long-term and stable expression.
3. The pseudo-type lentivirus is engineered so that it does not replicate and
is safe for most lab operations. Lentiviral platform is most frequently chosen
by customers. Other retrovirus system is also available. Details can be
provided if this platform is applicable to customer's cell line of interest.
Over-expressionOver-expressionOver-expressionOver-expression orororor knock-downknock-downknock-downknock-down
Over-expression or knock-down is one of the most widely used tools for
gene function research and disease target validation. Shanghai Genomics has
successfully generated hundreds of stable cell lines with either gene
over-expression or knock-down.
1. Over-expression cell lines: Many mammalian expression vectors or
lentiviral vectors are suitable for this purpose. Stable cell lines have been
generated in-house with pGL4; pcDNA3.1; pEF_Bos; pLenti-DEST vectors.
We may help customers to select a proper vector for their customized cell
line generation.
2. Knock-down cell lines: RNAi techniques are widely used for gene
knockdown applications. shRNA and miR-shRNA (a pseudo-type miRNA)
are major techniques used to generate SG's knockdown cell lines. Since
long-term suppression using pol III shRNAs can be problematic especially
for studies of genes that are critical to cell proliferation and differentiation,
we provide not only constitutive but also conditional knock-down. The table
below lists two major sets of lentiviral RNAi vectors used by SG for
knockdown cell line generation.
VectorVectorVectorVector
SetSetSetSet
SelectionSelectionSelectionSelection
MarkerMarkerMarkerMarker
ApproachApproachApproachApproach forforforfor
stablestablestablestable CellCellCellCell LineLineLineLine
generationgenerationgenerationgeneration
ApplicationApplicationApplicationApplication
pLKO.1
(Constitu
tive)
Puromyci
n
Drug selection
Knockdown of the target by
shRNA
EGFP Sort by FACS
Knockdown of the target by
shRNA
pSLIK
(Tet On)
Zeocin Drug selection
Knockdown of multiple targets
by miR-shRNAs
Neomyci
n
Drug selection
Knockdown of multiple targets
by miR-shRNAs
Hygromy
cin
Drug selection
Knockdown of multiple targets
by miR-shRNAs
CD4 Drug selection
Knockdown of multiple targets
by miR-shRNAs
Venus Sort by FACS
Knockdown of multiple targets
by miR-shRNAs
pSLIKpSLIKpSLIKpSLIK system:system:system:system: inducibleinducibleinducibleinducible lentivirallentivirallentivirallentiviral systemsystemsystemsystem (Tet-On)(Tet-On)(Tet-On)(Tet-On)
pLKO.1pLKO.1pLKO.1pLKO.1 system:system:system:system: constitutiveconstitutiveconstitutiveconstitutive lentivirallentivirallentivirallentiviral systemsystemsystemsystem
WorkflowWorkflowWorkflowWorkflow (using(using(using(using thethethethe lentivirallentivirallentivirallentiviral system)system)system)system)
There are many mature non-viral transfection products and many optimized
protocols available for stable cell line generation on the market. The
following flowchart shows the general workflow with the lentiviral infection
system. Generally, target expression will be investigated in established
stable cell lines. Mycoplasma testing and STR profiling are also done as part
of regular quality control to ensure mycoplasma free and identity between
master and established stable cell lines.
PricingPricingPricingPricing andandandand TimelineTimelineTimelineTimeline
The table below shows a rough price and timeline for customized stable cell
line generation using lentiviral infection. For different targets and cell lines,
or using nonviral transfection approach, please contact us for prices and
timelines.
ServicesServicesServicesServices PricePricePricePrice (in(in(in(in USD)USD)USD)USD) TimeTimeTimeTime linelinelineline
Probe design, construction
and validation
inquiry 5-7 weeks
pSLI
K
syste
m
Polyclonal cell line
generation and
validation
inquiry 9-10 weeks
Monoclonal cell
line generation and
validation
inquiry 13-14 weeks
pLK
O.1
Polyclonal cell line
generation and
validation
inquiry 8-9 weeks
Monoclonal cell
line generation and
validation
inquiry 12-13 weeks
ExamplesExamplesExamplesExamples
ExampleExampleExampleExample 1:1:1:1: miR-shRNA against luciferase was introduced into the
293T-Luc cell line by lentiviral infection and then sorted by FACS.
Constitutively expressed Luciferase was knocked down in the presence of
DOX.
ExampleExampleExampleExample 2:2:2:2: miR-shRNA against gene 1 or over-expression of gene 2 was
introduced into the master cell line by lentiviral infection and drug selection.
Significant knockdown or over-expression was observed by both RT-QPCR
and Western blot in the presence of DOX.
ContactContactContactContact Info:Info:Info:Info:
Linna Green
Project Management
Tel: 1-631-626-9181
Fax: 1-631-614-7828
Email: info@creative-biogene.com
(Creative(Creative(Creative(Creative DynamicsDynamicsDynamicsDynamics andandandand CDCDCDCD Bocsci)CDBocsci)CDBocsci)CDBocsci)CD,,,, Inc.Inc.Inc.Inc.
www.creative-biogene.com
45-16 Ramsey Road Shirley, NY,11967,USA

More Related Content

What's hot

Novartis BILS 2016
Novartis BILS 2016Novartis BILS 2016
Novartis BILS 2016GBX Events
 
CRISPR presentation extended Mouse Modeling
CRISPR presentation extended Mouse ModelingCRISPR presentation extended Mouse Modeling
CRISPR presentation extended Mouse ModelingTristan Kempston
 
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...Merck Life Sciences
 
Avacta Life Sciences Affimers Presentation Global Protein Engineering Summit ...
Avacta Life Sciences Affimers Presentation Global Protein Engineering Summit ...Avacta Life Sciences Affimers Presentation Global Protein Engineering Summit ...
Avacta Life Sciences Affimers Presentation Global Protein Engineering Summit ...AvactaLifeSciences
 
Clinical molecular diagnostics for drug guidance
Clinical molecular diagnostics for drug guidanceClinical molecular diagnostics for drug guidance
Clinical molecular diagnostics for drug guidanceNikesh Shah
 
Towards Precision Medicine: Tute Genomics, a cloud-based application for anal...
Towards Precision Medicine: Tute Genomics, a cloud-based application for anal...Towards Precision Medicine: Tute Genomics, a cloud-based application for anal...
Towards Precision Medicine: Tute Genomics, a cloud-based application for anal...Reid Robison
 
Process improvement delivered by a high efficiency, automated single cell clo...
Process improvement delivered by a high efficiency, automated single cell clo...Process improvement delivered by a high efficiency, automated single cell clo...
Process improvement delivered by a high efficiency, automated single cell clo...IanTaylor50
 
CRISPR: what it is, and why it is having a profound impact on human health
CRISPR: what it is, and why it is having a profound impact on human healthCRISPR: what it is, and why it is having a profound impact on human health
CRISPR: what it is, and why it is having a profound impact on human healthPistoia Alliance
 
Unveiling the Potential of your AAV Gene Therapy: Orthogonal methods to under...
Unveiling the Potential of your AAV Gene Therapy: Orthogonal methods to under...Unveiling the Potential of your AAV Gene Therapy: Orthogonal methods to under...
Unveiling the Potential of your AAV Gene Therapy: Orthogonal methods to under...Merck Life Sciences
 
The Trans-NIH RNAi Initiative : Informatics
The Trans-NIH RNAi Initiative: InformaticsThe Trans-NIH RNAi Initiative: Informatics
The Trans-NIH RNAi Initiative : InformaticsRajarshi Guha
 
CRISPR Crops--a talk by Sophien Kamoun at Science Portal BD
CRISPR Crops--a talk by Sophien Kamoun at Science Portal BDCRISPR Crops--a talk by Sophien Kamoun at Science Portal BD
CRISPR Crops--a talk by Sophien Kamoun at Science Portal BDSophien Kamoun
 
Translating Genomes | Personalizing Medicine
Translating Genomes | Personalizing MedicineTranslating Genomes | Personalizing Medicine
Translating Genomes | Personalizing MedicineCandy Smellie
 
Production and purification of Viral vectors for gene and cell therapy appli...
Production and purification of  Viral vectors for gene and cell therapy appli...Production and purification of  Viral vectors for gene and cell therapy appli...
Production and purification of Viral vectors for gene and cell therapy appli...Dr. Priyabrata Pattnaik
 
Improved Platforms for CHO Cell Line Development
Improved Platforms for CHO Cell Line Development  Improved Platforms for CHO Cell Line Development
Improved Platforms for CHO Cell Line Development Merck Life Sciences
 
I psc and stem cell 2013
I psc and stem cell 2013I psc and stem cell 2013
I psc and stem cell 2013Elsa von Licy
 
In Vitro Analog of the Primitive Streak (ANIMATED)
In Vitro Analog of the Primitive Streak (ANIMATED)In Vitro Analog of the Primitive Streak (ANIMATED)
In Vitro Analog of the Primitive Streak (ANIMATED)Nikolay Turovets
 
CRISPR-Revolutionary Genome editing tools for Plants.....
CRISPR-Revolutionary Genome editing tools for Plants.....CRISPR-Revolutionary Genome editing tools for Plants.....
CRISPR-Revolutionary Genome editing tools for Plants.....BHU,Varanasi, INDIA
 
Institute of Learning in Retirement - Miami University (Ohio)
Institute of Learning in Retirement - Miami University (Ohio)Institute of Learning in Retirement - Miami University (Ohio)
Institute of Learning in Retirement - Miami University (Ohio)Andor Kiss
 

What's hot (20)

Novartis BILS 2016
Novartis BILS 2016Novartis BILS 2016
Novartis BILS 2016
 
CRISPR presentation extended Mouse Modeling
CRISPR presentation extended Mouse ModelingCRISPR presentation extended Mouse Modeling
CRISPR presentation extended Mouse Modeling
 
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...
 
Avacta Life Sciences Affimers Presentation Global Protein Engineering Summit ...
Avacta Life Sciences Affimers Presentation Global Protein Engineering Summit ...Avacta Life Sciences Affimers Presentation Global Protein Engineering Summit ...
Avacta Life Sciences Affimers Presentation Global Protein Engineering Summit ...
 
Clinical molecular diagnostics for drug guidance
Clinical molecular diagnostics for drug guidanceClinical molecular diagnostics for drug guidance
Clinical molecular diagnostics for drug guidance
 
Towards Precision Medicine: Tute Genomics, a cloud-based application for anal...
Towards Precision Medicine: Tute Genomics, a cloud-based application for anal...Towards Precision Medicine: Tute Genomics, a cloud-based application for anal...
Towards Precision Medicine: Tute Genomics, a cloud-based application for anal...
 
Process improvement delivered by a high efficiency, automated single cell clo...
Process improvement delivered by a high efficiency, automated single cell clo...Process improvement delivered by a high efficiency, automated single cell clo...
Process improvement delivered by a high efficiency, automated single cell clo...
 
CRISPR: what it is, and why it is having a profound impact on human health
CRISPR: what it is, and why it is having a profound impact on human healthCRISPR: what it is, and why it is having a profound impact on human health
CRISPR: what it is, and why it is having a profound impact on human health
 
Unveiling the Potential of your AAV Gene Therapy: Orthogonal methods to under...
Unveiling the Potential of your AAV Gene Therapy: Orthogonal methods to under...Unveiling the Potential of your AAV Gene Therapy: Orthogonal methods to under...
Unveiling the Potential of your AAV Gene Therapy: Orthogonal methods to under...
 
The Trans-NIH RNAi Initiative : Informatics
The Trans-NIH RNAi Initiative: InformaticsThe Trans-NIH RNAi Initiative: Informatics
The Trans-NIH RNAi Initiative : Informatics
 
CRISPR Crops--a talk by Sophien Kamoun at Science Portal BD
CRISPR Crops--a talk by Sophien Kamoun at Science Portal BDCRISPR Crops--a talk by Sophien Kamoun at Science Portal BD
CRISPR Crops--a talk by Sophien Kamoun at Science Portal BD
 
Translating Genomes | Personalizing Medicine
Translating Genomes | Personalizing MedicineTranslating Genomes | Personalizing Medicine
Translating Genomes | Personalizing Medicine
 
Production and purification of Viral vectors for gene and cell therapy appli...
Production and purification of  Viral vectors for gene and cell therapy appli...Production and purification of  Viral vectors for gene and cell therapy appli...
Production and purification of Viral vectors for gene and cell therapy appli...
 
Improved Platforms for CHO Cell Line Development
Improved Platforms for CHO Cell Line Development  Improved Platforms for CHO Cell Line Development
Improved Platforms for CHO Cell Line Development
 
I psc and stem cell 2013
I psc and stem cell 2013I psc and stem cell 2013
I psc and stem cell 2013
 
Crispr cas
Crispr casCrispr cas
Crispr cas
 
In Vitro Analog of the Primitive Streak (ANIMATED)
In Vitro Analog of the Primitive Streak (ANIMATED)In Vitro Analog of the Primitive Streak (ANIMATED)
In Vitro Analog of the Primitive Streak (ANIMATED)
 
CRISPR-Revolutionary Genome editing tools for Plants.....
CRISPR-Revolutionary Genome editing tools for Plants.....CRISPR-Revolutionary Genome editing tools for Plants.....
CRISPR-Revolutionary Genome editing tools for Plants.....
 
Institute of Learning in Retirement - Miami University (Ohio)
Institute of Learning in Retirement - Miami University (Ohio)Institute of Learning in Retirement - Miami University (Ohio)
Institute of Learning in Retirement - Miami University (Ohio)
 
webconferences.com/nihoba/2003sep17
webconferences.com/nihoba/2003sep17webconferences.com/nihoba/2003sep17
webconferences.com/nihoba/2003sep17
 

Similar to Customized stable cell line services

Reporter stem cells generation
Reporter stem cells generationReporter stem cells generation
Reporter stem cells generationCreative Biogene
 
Cell Authentication By STR Profiling
Cell Authentication By STR ProfilingCell Authentication By STR Profiling
Cell Authentication By STR ProfilingCreative-Bioarray
 
Cell authentication by str profile
Cell authentication by str profileCell authentication by str profile
Cell authentication by str profileBennie George
 
short tandem repeats profile
short tandem repeats profileshort tandem repeats profile
short tandem repeats profileBennie George
 
Reporter stem cells generation
Reporter stem cells generation Reporter stem cells generation
Reporter stem cells generation Stella Evelyn
 
Standardization of human stem cell pluripotency using bioinformatics presenta...
Standardization of human stem cell pluripotency using bioinformatics presenta...Standardization of human stem cell pluripotency using bioinformatics presenta...
Standardization of human stem cell pluripotency using bioinformatics presenta...بدر العلوان
 
Reporter stem cells generation
Reporter stem cells generationReporter stem cells generation
Reporter stem cells generationCreative Biogene
 
Chimeric Antigen Receptor Technology.pdf
Chimeric Antigen Receptor Technology.pdfChimeric Antigen Receptor Technology.pdf
Chimeric Antigen Receptor Technology.pdfCandy Swift
 
LanglaisC_2007_Systematic approach to protein experssion in cell free systems
LanglaisC_2007_Systematic approach to protein experssion in cell free systemsLanglaisC_2007_Systematic approach to protein experssion in cell free systems
LanglaisC_2007_Systematic approach to protein experssion in cell free systemsClaudia Langlais
 
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...MilliporeSigma
 
Integrating arrays and RNA-Seq
Integrating arrays and RNA-Seq Integrating arrays and RNA-Seq
Integrating arrays and RNA-Seq Affymetrix
 
Mammalian & Bacterial Expression
Mammalian & Bacterial ExpressionMammalian & Bacterial Expression
Mammalian & Bacterial ExpressionJesús C. Morales
 
Accelerating Usp
Accelerating UspAccelerating Usp
Accelerating UspSMEbiotech
 
FFPE Applications Solutions brochure
FFPE Applications Solutions brochureFFPE Applications Solutions brochure
FFPE Applications Solutions brochureAffymetrix
 
Stable knockout cell line generation
Stable knockout cell line generationStable knockout cell line generation
Stable knockout cell line generationCreative Biogene
 
Stable knockout cell line generation
Stable knockout cell line generationStable knockout cell line generation
Stable knockout cell line generationStella Evelyn
 
Spotlight on ipsc reprogramming
Spotlight on ipsc reprogrammingSpotlight on ipsc reprogramming
Spotlight on ipsc reprogrammingCreative-Bioarray
 

Similar to Customized stable cell line services (20)

Reporter stem cells generation
Reporter stem cells generationReporter stem cells generation
Reporter stem cells generation
 
Cell Authentication By STR Profiling
Cell Authentication By STR ProfilingCell Authentication By STR Profiling
Cell Authentication By STR Profiling
 
Cell authentication by str profile
Cell authentication by str profileCell authentication by str profile
Cell authentication by str profile
 
short tandem repeats profile
short tandem repeats profileshort tandem repeats profile
short tandem repeats profile
 
Reporter stem cells generation
Reporter stem cells generation Reporter stem cells generation
Reporter stem cells generation
 
Standardization of human stem cell pluripotency using bioinformatics presenta...
Standardization of human stem cell pluripotency using bioinformatics presenta...Standardization of human stem cell pluripotency using bioinformatics presenta...
Standardization of human stem cell pluripotency using bioinformatics presenta...
 
Reporter stem cells generation
Reporter stem cells generationReporter stem cells generation
Reporter stem cells generation
 
Chimeric Antigen Receptor Technology.pdf
Chimeric Antigen Receptor Technology.pdfChimeric Antigen Receptor Technology.pdf
Chimeric Antigen Receptor Technology.pdf
 
LanglaisC_2007_Systematic approach to protein experssion in cell free systems
LanglaisC_2007_Systematic approach to protein experssion in cell free systemsLanglaisC_2007_Systematic approach to protein experssion in cell free systems
LanglaisC_2007_Systematic approach to protein experssion in cell free systems
 
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...
Releasing Your AAV Therapy with Confidence: Regulatory Considerations and Key...
 
Integrating arrays and RNA-Seq
Integrating arrays and RNA-Seq Integrating arrays and RNA-Seq
Integrating arrays and RNA-Seq
 
Mammalian & Bacterial Expression
Mammalian & Bacterial ExpressionMammalian & Bacterial Expression
Mammalian & Bacterial Expression
 
Accelerating Usp
Accelerating UspAccelerating Usp
Accelerating Usp
 
Sh rna 2013
Sh rna 2013Sh rna 2013
Sh rna 2013
 
MSU Transgenic and Genome Editing Facility
MSU Transgenic and Genome Editing FacilityMSU Transgenic and Genome Editing Facility
MSU Transgenic and Genome Editing Facility
 
FFPE Applications Solutions brochure
FFPE Applications Solutions brochureFFPE Applications Solutions brochure
FFPE Applications Solutions brochure
 
Stable knockout cell line generation
Stable knockout cell line generationStable knockout cell line generation
Stable knockout cell line generation
 
Stable knockout cell line generation
Stable knockout cell line generationStable knockout cell line generation
Stable knockout cell line generation
 
iPSC-derived Cardiomyocytes
iPSC-derived CardiomyocytesiPSC-derived Cardiomyocytes
iPSC-derived Cardiomyocytes
 
Spotlight on ipsc reprogramming
Spotlight on ipsc reprogrammingSpotlight on ipsc reprogramming
Spotlight on ipsc reprogramming
 

Recently uploaded

Linked Data in Production: Moving Beyond Ontologies
Linked Data in Production: Moving Beyond OntologiesLinked Data in Production: Moving Beyond Ontologies
Linked Data in Production: Moving Beyond OntologiesDavid Newbury
 
Designing A Time bound resource download URL
Designing A Time bound resource download URLDesigning A Time bound resource download URL
Designing A Time bound resource download URLRuncy Oommen
 
Using IESVE for Loads, Sizing and Heat Pump Modeling to Achieve Decarbonization
Using IESVE for Loads, Sizing and Heat Pump Modeling to Achieve DecarbonizationUsing IESVE for Loads, Sizing and Heat Pump Modeling to Achieve Decarbonization
Using IESVE for Loads, Sizing and Heat Pump Modeling to Achieve DecarbonizationIES VE
 
Cloud Revolution: Exploring the New Wave of Serverless Spatial Data
Cloud Revolution: Exploring the New Wave of Serverless Spatial DataCloud Revolution: Exploring the New Wave of Serverless Spatial Data
Cloud Revolution: Exploring the New Wave of Serverless Spatial DataSafe Software
 
UiPath Community: AI for UiPath Automation Developers
UiPath Community: AI for UiPath Automation DevelopersUiPath Community: AI for UiPath Automation Developers
UiPath Community: AI for UiPath Automation DevelopersUiPathCommunity
 
NIST Cybersecurity Framework (CSF) 2.0 Workshop
NIST Cybersecurity Framework (CSF) 2.0 WorkshopNIST Cybersecurity Framework (CSF) 2.0 Workshop
NIST Cybersecurity Framework (CSF) 2.0 WorkshopBachir Benyammi
 
Meet the new FSP 3000 M-Flex800™
Meet the new FSP 3000 M-Flex800™Meet the new FSP 3000 M-Flex800™
Meet the new FSP 3000 M-Flex800™Adtran
 
Introduction to Quantum Computing
Introduction to Quantum ComputingIntroduction to Quantum Computing
Introduction to Quantum ComputingGDSC PJATK
 
IaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdf
IaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdfIaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdf
IaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdfDaniel Santiago Silva Capera
 
Anypoint Code Builder , Google Pub sub connector and MuleSoft RPA
Anypoint Code Builder , Google Pub sub connector and MuleSoft RPAAnypoint Code Builder , Google Pub sub connector and MuleSoft RPA
Anypoint Code Builder , Google Pub sub connector and MuleSoft RPAshyamraj55
 
Do we need a new standard for visualizing the invisible?
Do we need a new standard for visualizing the invisible?Do we need a new standard for visualizing the invisible?
Do we need a new standard for visualizing the invisible?SANGHEE SHIN
 
Artificial Intelligence & SEO Trends for 2024
Artificial Intelligence & SEO Trends for 2024Artificial Intelligence & SEO Trends for 2024
Artificial Intelligence & SEO Trends for 2024D Cloud Solutions
 
Babel Compiler - Transforming JavaScript for All Browsers.pptx
Babel Compiler - Transforming JavaScript for All Browsers.pptxBabel Compiler - Transforming JavaScript for All Browsers.pptx
Babel Compiler - Transforming JavaScript for All Browsers.pptxYounusS2
 
UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...
UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...
UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...UbiTrack UK
 
Cybersecurity Workshop #1.pptx
Cybersecurity Workshop #1.pptxCybersecurity Workshop #1.pptx
Cybersecurity Workshop #1.pptxGDSC PJATK
 
Videogame localization & technology_ how to enhance the power of translation.pdf
Videogame localization & technology_ how to enhance the power of translation.pdfVideogame localization & technology_ how to enhance the power of translation.pdf
Videogame localization & technology_ how to enhance the power of translation.pdfinfogdgmi
 
20200723_insight_release_plan_v6.pdf20200723_insight_release_plan_v6.pdf
20200723_insight_release_plan_v6.pdf20200723_insight_release_plan_v6.pdf20200723_insight_release_plan_v6.pdf20200723_insight_release_plan_v6.pdf
20200723_insight_release_plan_v6.pdf20200723_insight_release_plan_v6.pdfJamie (Taka) Wang
 
UiPath Platform: The Backend Engine Powering Your Automation - Session 1
UiPath Platform: The Backend Engine Powering Your Automation - Session 1UiPath Platform: The Backend Engine Powering Your Automation - Session 1
UiPath Platform: The Backend Engine Powering Your Automation - Session 1DianaGray10
 
Salesforce Miami User Group Event - 1st Quarter 2024
Salesforce Miami User Group Event - 1st Quarter 2024Salesforce Miami User Group Event - 1st Quarter 2024
Salesforce Miami User Group Event - 1st Quarter 2024SkyPlanner
 
UiPath Solutions Management Preview - Northern CA Chapter - March 22.pdf
UiPath Solutions Management Preview - Northern CA Chapter - March 22.pdfUiPath Solutions Management Preview - Northern CA Chapter - March 22.pdf
UiPath Solutions Management Preview - Northern CA Chapter - March 22.pdfDianaGray10
 

Recently uploaded (20)

Linked Data in Production: Moving Beyond Ontologies
Linked Data in Production: Moving Beyond OntologiesLinked Data in Production: Moving Beyond Ontologies
Linked Data in Production: Moving Beyond Ontologies
 
Designing A Time bound resource download URL
Designing A Time bound resource download URLDesigning A Time bound resource download URL
Designing A Time bound resource download URL
 
Using IESVE for Loads, Sizing and Heat Pump Modeling to Achieve Decarbonization
Using IESVE for Loads, Sizing and Heat Pump Modeling to Achieve DecarbonizationUsing IESVE for Loads, Sizing and Heat Pump Modeling to Achieve Decarbonization
Using IESVE for Loads, Sizing and Heat Pump Modeling to Achieve Decarbonization
 
Cloud Revolution: Exploring the New Wave of Serverless Spatial Data
Cloud Revolution: Exploring the New Wave of Serverless Spatial DataCloud Revolution: Exploring the New Wave of Serverless Spatial Data
Cloud Revolution: Exploring the New Wave of Serverless Spatial Data
 
UiPath Community: AI for UiPath Automation Developers
UiPath Community: AI for UiPath Automation DevelopersUiPath Community: AI for UiPath Automation Developers
UiPath Community: AI for UiPath Automation Developers
 
NIST Cybersecurity Framework (CSF) 2.0 Workshop
NIST Cybersecurity Framework (CSF) 2.0 WorkshopNIST Cybersecurity Framework (CSF) 2.0 Workshop
NIST Cybersecurity Framework (CSF) 2.0 Workshop
 
Meet the new FSP 3000 M-Flex800™
Meet the new FSP 3000 M-Flex800™Meet the new FSP 3000 M-Flex800™
Meet the new FSP 3000 M-Flex800™
 
Introduction to Quantum Computing
Introduction to Quantum ComputingIntroduction to Quantum Computing
Introduction to Quantum Computing
 
IaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdf
IaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdfIaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdf
IaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdf
 
Anypoint Code Builder , Google Pub sub connector and MuleSoft RPA
Anypoint Code Builder , Google Pub sub connector and MuleSoft RPAAnypoint Code Builder , Google Pub sub connector and MuleSoft RPA
Anypoint Code Builder , Google Pub sub connector and MuleSoft RPA
 
Do we need a new standard for visualizing the invisible?
Do we need a new standard for visualizing the invisible?Do we need a new standard for visualizing the invisible?
Do we need a new standard for visualizing the invisible?
 
Artificial Intelligence & SEO Trends for 2024
Artificial Intelligence & SEO Trends for 2024Artificial Intelligence & SEO Trends for 2024
Artificial Intelligence & SEO Trends for 2024
 
Babel Compiler - Transforming JavaScript for All Browsers.pptx
Babel Compiler - Transforming JavaScript for All Browsers.pptxBabel Compiler - Transforming JavaScript for All Browsers.pptx
Babel Compiler - Transforming JavaScript for All Browsers.pptx
 
UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...
UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...
UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...
 
Cybersecurity Workshop #1.pptx
Cybersecurity Workshop #1.pptxCybersecurity Workshop #1.pptx
Cybersecurity Workshop #1.pptx
 
Videogame localization & technology_ how to enhance the power of translation.pdf
Videogame localization & technology_ how to enhance the power of translation.pdfVideogame localization & technology_ how to enhance the power of translation.pdf
Videogame localization & technology_ how to enhance the power of translation.pdf
 
20200723_insight_release_plan_v6.pdf20200723_insight_release_plan_v6.pdf
20200723_insight_release_plan_v6.pdf20200723_insight_release_plan_v6.pdf20200723_insight_release_plan_v6.pdf20200723_insight_release_plan_v6.pdf
20200723_insight_release_plan_v6.pdf20200723_insight_release_plan_v6.pdf
 
UiPath Platform: The Backend Engine Powering Your Automation - Session 1
UiPath Platform: The Backend Engine Powering Your Automation - Session 1UiPath Platform: The Backend Engine Powering Your Automation - Session 1
UiPath Platform: The Backend Engine Powering Your Automation - Session 1
 
Salesforce Miami User Group Event - 1st Quarter 2024
Salesforce Miami User Group Event - 1st Quarter 2024Salesforce Miami User Group Event - 1st Quarter 2024
Salesforce Miami User Group Event - 1st Quarter 2024
 
UiPath Solutions Management Preview - Northern CA Chapter - March 22.pdf
UiPath Solutions Management Preview - Northern CA Chapter - March 22.pdfUiPath Solutions Management Preview - Northern CA Chapter - March 22.pdf
UiPath Solutions Management Preview - Northern CA Chapter - March 22.pdf
 

Customized stable cell line services

  • 1. CustomizedCustomizedCustomizedCustomized ServicesServicesServicesServices ForForForFor StableStableStableStable Knock-DownKnock-DownKnock-DownKnock-Down orororor Over-ExpressionOver-ExpressionOver-ExpressionOver-Expression CellCellCellCell LineLineLineLine GenerationGenerationGenerationGeneration Stable cell lines with specific gene over-expression or knock-down are very helpful in gene function analysis, target discovery, target validation, assay development, and compound screening. However, generation of stable cell lines is a time-consuming and expensive process. In the last 4 years, Shanghai Genomics has established a team of experts for stable cell line generation. We have successfully generated hundreds of stable cell lines to meet the need of our customers. MethodMethodMethodMethod Shanghai Genomics typically uses non-viral (liposome) and viral (lentivirus) platforms for stable cell line generation. For some cell lines, like HEK293 and HeLa, the liposome method can achieve high transfection efficiency. Compared to liposomal or other chemical vehicles, a lentiviral approach has three distinct advantages: 1. The pseudotype virus has a broad range of host cell lines and is able to infect both dividing and non-dividing cells, making it especially suitable for some differentiated cell lines such as neurons, macrophages, hematopoietic stem cells, retinal photoreceptors and muscle and liver cells. 2. The virally encoded integrase greatly facilitates exogenous gene integration that ensures long-term and stable expression. 3. The pseudo-type lentivirus is engineered so that it does not replicate and is safe for most lab operations. Lentiviral platform is most frequently chosen by customers. Other retrovirus system is also available. Details can be provided if this platform is applicable to customer's cell line of interest. Over-expressionOver-expressionOver-expressionOver-expression orororor knock-downknock-downknock-downknock-down Over-expression or knock-down is one of the most widely used tools for gene function research and disease target validation. Shanghai Genomics has
  • 2. successfully generated hundreds of stable cell lines with either gene over-expression or knock-down. 1. Over-expression cell lines: Many mammalian expression vectors or lentiviral vectors are suitable for this purpose. Stable cell lines have been generated in-house with pGL4; pcDNA3.1; pEF_Bos; pLenti-DEST vectors. We may help customers to select a proper vector for their customized cell line generation. 2. Knock-down cell lines: RNAi techniques are widely used for gene knockdown applications. shRNA and miR-shRNA (a pseudo-type miRNA) are major techniques used to generate SG's knockdown cell lines. Since long-term suppression using pol III shRNAs can be problematic especially for studies of genes that are critical to cell proliferation and differentiation, we provide not only constitutive but also conditional knock-down. The table below lists two major sets of lentiviral RNAi vectors used by SG for knockdown cell line generation. VectorVectorVectorVector SetSetSetSet SelectionSelectionSelectionSelection MarkerMarkerMarkerMarker ApproachApproachApproachApproach forforforfor stablestablestablestable CellCellCellCell LineLineLineLine generationgenerationgenerationgeneration ApplicationApplicationApplicationApplication pLKO.1 (Constitu tive) Puromyci n Drug selection Knockdown of the target by shRNA EGFP Sort by FACS Knockdown of the target by shRNA pSLIK (Tet On) Zeocin Drug selection Knockdown of multiple targets by miR-shRNAs Neomyci n Drug selection Knockdown of multiple targets by miR-shRNAs Hygromy cin Drug selection Knockdown of multiple targets by miR-shRNAs CD4 Drug selection Knockdown of multiple targets by miR-shRNAs Venus Sort by FACS Knockdown of multiple targets by miR-shRNAs
  • 3. pSLIKpSLIKpSLIKpSLIK system:system:system:system: inducibleinducibleinducibleinducible lentivirallentivirallentivirallentiviral systemsystemsystemsystem (Tet-On)(Tet-On)(Tet-On)(Tet-On) pLKO.1pLKO.1pLKO.1pLKO.1 system:system:system:system: constitutiveconstitutiveconstitutiveconstitutive lentivirallentivirallentivirallentiviral systemsystemsystemsystem WorkflowWorkflowWorkflowWorkflow (using(using(using(using thethethethe lentivirallentivirallentivirallentiviral system)system)system)system) There are many mature non-viral transfection products and many optimized protocols available for stable cell line generation on the market. The following flowchart shows the general workflow with the lentiviral infection system. Generally, target expression will be investigated in established stable cell lines. Mycoplasma testing and STR profiling are also done as part of regular quality control to ensure mycoplasma free and identity between master and established stable cell lines.
  • 4. PricingPricingPricingPricing andandandand TimelineTimelineTimelineTimeline The table below shows a rough price and timeline for customized stable cell line generation using lentiviral infection. For different targets and cell lines, or using nonviral transfection approach, please contact us for prices and timelines. ServicesServicesServicesServices PricePricePricePrice (in(in(in(in USD)USD)USD)USD) TimeTimeTimeTime linelinelineline Probe design, construction and validation inquiry 5-7 weeks pSLI K syste m Polyclonal cell line generation and validation inquiry 9-10 weeks Monoclonal cell line generation and validation inquiry 13-14 weeks pLK O.1 Polyclonal cell line generation and validation inquiry 8-9 weeks Monoclonal cell line generation and validation inquiry 12-13 weeks ExamplesExamplesExamplesExamples
  • 5. ExampleExampleExampleExample 1:1:1:1: miR-shRNA against luciferase was introduced into the 293T-Luc cell line by lentiviral infection and then sorted by FACS. Constitutively expressed Luciferase was knocked down in the presence of DOX. ExampleExampleExampleExample 2:2:2:2: miR-shRNA against gene 1 or over-expression of gene 2 was introduced into the master cell line by lentiviral infection and drug selection. Significant knockdown or over-expression was observed by both RT-QPCR and Western blot in the presence of DOX. ContactContactContactContact Info:Info:Info:Info: Linna Green Project Management Tel: 1-631-626-9181 Fax: 1-631-614-7828 Email: info@creative-biogene.com (Creative(Creative(Creative(Creative DynamicsDynamicsDynamicsDynamics andandandand CDCDCDCD Bocsci)CDBocsci)CDBocsci)CDBocsci)CD,,,, Inc.Inc.Inc.Inc. www.creative-biogene.com 45-16 Ramsey Road Shirley, NY,11967,USA