CRISPR/Cas9 is a genome editing technique that allows for highly specific modification of DNA. It involves a Cas9 protein guiding a customized RNA to a target location in the genome to cut DNA. Scientists adapted the natural CRISPR immune system found in bacteria for genome editing. CRISPR/Cas9 holds promise for treating genetic diseases and developing crops but also raises ethical concerns when applied to human germline cells due to heritable effects. Researchers continue improving targeting specificity and developing new Cas proteins for additional applications in genome editing and gene regulation.
CRISPR (clustered regularly interspaced short palindromic repeats) is a family of DNA sequences found within the genomes of prokaryotic organisms such as bacteria and archaea. These sequences are derived from DNA fragments of bacteriophages that have previously infected the prokaryote and are used to detect and destroy DNA from similar phages during subsequent infections. Hence these sequences play a key role in the antiviral defense system of prokaryotes.
Cas9 (CRISPR-associated protein 9) is an enzyme that uses CRISPR sequences as a guide to recognize and cleave specific strands of DNA that are complementary to the CRISPR sequence. Cas9 enzymes together with CRISPR sequences form the basis of a technology known as CRISPR-Cas9 that can be used to edit genes within organisms.This editing process has a wide variety of applications including basic biological research, development of biotechnology products, and treatment of diseases.
The CRISPR-Cas system is a prokaryotic immune system that confers resistance to foreign genetic elements such as those present within plasmids and phages that provides a form of acquired immunity. RNA harboring the spacer sequence helps Cas (CRISPR-associated) proteins recognize and cut foreign pathogenic DNA. Other RNA-guided Cas proteins cut foreign RNA. CRISPR are found in approximately 50% of sequenced bacterial genomes and nearly 90% of sequenced archaea.
Genome editing with the CRISPR-Cas9 system has become one of the major tools in modern biotechnology. This slide share discusses the fundamentals in a simple, easy to understand format.
i explained about basics of genome engineering and crispr system.
CRISPR will change the world and it is just the beginning, are you ready to meet the future? you think its great and beautiful or.....?
please give your feedback to my email
pooyanaghshbandi@yahoo.com
i am starting to write a critical and fantastic review article about CRISPR, if you are interested to join please contact me.
CRISPR (clustered regularly interspaced short palindromic repeats) is a family of DNA sequences found within the genomes of prokaryotic organisms such as bacteria and archaea. These sequences are derived from DNA fragments of bacteriophages that have previously infected the prokaryote and are used to detect and destroy DNA from similar phages during subsequent infections. Hence these sequences play a key role in the antiviral defense system of prokaryotes.
Cas9 (CRISPR-associated protein 9) is an enzyme that uses CRISPR sequences as a guide to recognize and cleave specific strands of DNA that are complementary to the CRISPR sequence. Cas9 enzymes together with CRISPR sequences form the basis of a technology known as CRISPR-Cas9 that can be used to edit genes within organisms.This editing process has a wide variety of applications including basic biological research, development of biotechnology products, and treatment of diseases.
The CRISPR-Cas system is a prokaryotic immune system that confers resistance to foreign genetic elements such as those present within plasmids and phages that provides a form of acquired immunity. RNA harboring the spacer sequence helps Cas (CRISPR-associated) proteins recognize and cut foreign pathogenic DNA. Other RNA-guided Cas proteins cut foreign RNA. CRISPR are found in approximately 50% of sequenced bacterial genomes and nearly 90% of sequenced archaea.
Genome editing with the CRISPR-Cas9 system has become one of the major tools in modern biotechnology. This slide share discusses the fundamentals in a simple, easy to understand format.
i explained about basics of genome engineering and crispr system.
CRISPR will change the world and it is just the beginning, are you ready to meet the future? you think its great and beautiful or.....?
please give your feedback to my email
pooyanaghshbandi@yahoo.com
i am starting to write a critical and fantastic review article about CRISPR, if you are interested to join please contact me.
a brief description on the new emerging genome editing technology CRISPR-Cas9. this technique is making its place stronger and stronger day by day. and impossible things can be possible by this technique. and some main and famous names who discovered this technique.
CRISPR-Cas9 is a genome editing tool that is creating a buzz in the science world. It is faster, cheaper and more accurate than previous techniques of editing DNA and has a wide range of potential applications.
Have you considered that protein over-expression or inefficient mRNA knockdown may be masking physiological effects in your assays? Increasingly scientists are moving to endogenous gene-editing to characterise the function of their genes of interest.
Dr Chris Thorne from Cambridge Biotech Horizon Discovery discusses the ground breaking gene-editing technology CRISPR. The simplicity of experimental design has led to rapid adoption of the technology across the scientific community. However, challenges remain.
This Slidedeck focuses specifically on implementing CRISPR experiments, and explore a number of key considerations crucial to maximising chances of targeting success, whether your goal is to generate a knock-out or a knock-in. Chris also takes a look at some of the alternative uses of CRISPR, including sgRNA genome wide synthetic lethality screens.
The slides aim to support those researchers either planning to or already using CRISPR gene-editing in their lab. Horizon Discovery have also recently launched a program aimed specifically at academic cell biologists to promote the adoption of CRISPR by offering FREE CRISPR Reagents for knock-out cell line generation - more information available here. http://www.horizondiscovery.com/what-we-do/discovery-toolbox/genassist-crispr--raav-genome-editing-tools
A simple version of the CRISPR/Cas system, CRISPR/Cas9, has been modified to edit genomes. By delivering the Cas9 nuclease complexed with a synthetic guide RNA (gRNA) into a cell, the cell's genome can be cut at a desired location, allowing existing genes to be removed and/or new ones added.
An Introduction to Crispr Genome Editing
Crispr cas: A new tool of genome editing
CRISPRs (Clustered Regularly Interspaced Short Palindromic Repeats) are part of an adaptive defense mechanism in bacteria and archaea. Use of the CRISPR/Cas9 system for genome editing has been a major technological breakthrough, making genome modification in cells or organisms fast, more efficient, and much more robust than previous genome editing methods. Single guide RNAs (sgRNAs) or guide RNAs (gRNAs) direct and activate the Cas9 endonuclease at a specific genomic sequence. Cas9 then cleaves the target DNA, making it available for repair by the non-homologous end joining (NHEJ) system or for creating an insertion site for exogenous donor DNA by homologous recombination.
a brief description on the new emerging genome editing technology CRISPR-Cas9. this technique is making its place stronger and stronger day by day. and impossible things can be possible by this technique. and some main and famous names who discovered this technique.
CRISPR-Cas9 is a genome editing tool that is creating a buzz in the science world. It is faster, cheaper and more accurate than previous techniques of editing DNA and has a wide range of potential applications.
Have you considered that protein over-expression or inefficient mRNA knockdown may be masking physiological effects in your assays? Increasingly scientists are moving to endogenous gene-editing to characterise the function of their genes of interest.
Dr Chris Thorne from Cambridge Biotech Horizon Discovery discusses the ground breaking gene-editing technology CRISPR. The simplicity of experimental design has led to rapid adoption of the technology across the scientific community. However, challenges remain.
This Slidedeck focuses specifically on implementing CRISPR experiments, and explore a number of key considerations crucial to maximising chances of targeting success, whether your goal is to generate a knock-out or a knock-in. Chris also takes a look at some of the alternative uses of CRISPR, including sgRNA genome wide synthetic lethality screens.
The slides aim to support those researchers either planning to or already using CRISPR gene-editing in their lab. Horizon Discovery have also recently launched a program aimed specifically at academic cell biologists to promote the adoption of CRISPR by offering FREE CRISPR Reagents for knock-out cell line generation - more information available here. http://www.horizondiscovery.com/what-we-do/discovery-toolbox/genassist-crispr--raav-genome-editing-tools
A simple version of the CRISPR/Cas system, CRISPR/Cas9, has been modified to edit genomes. By delivering the Cas9 nuclease complexed with a synthetic guide RNA (gRNA) into a cell, the cell's genome can be cut at a desired location, allowing existing genes to be removed and/or new ones added.
An Introduction to Crispr Genome Editing
Crispr cas: A new tool of genome editing
CRISPRs (Clustered Regularly Interspaced Short Palindromic Repeats) are part of an adaptive defense mechanism in bacteria and archaea. Use of the CRISPR/Cas9 system for genome editing has been a major technological breakthrough, making genome modification in cells or organisms fast, more efficient, and much more robust than previous genome editing methods. Single guide RNAs (sgRNAs) or guide RNAs (gRNAs) direct and activate the Cas9 endonuclease at a specific genomic sequence. Cas9 then cleaves the target DNA, making it available for repair by the non-homologous end joining (NHEJ) system or for creating an insertion site for exogenous donor DNA by homologous recombination.
CRISPR is one of the mind blowing discovery which completely change the science of microorganisms. It is am efficient tool for genome editing and make the scientist enable to treat disease. The vast application of CRISPR technology covered almost all every aspect of life ranging from individual life to commercial aspect.
Purpose:
The purpose of this webinar is to develop creative scientific thinking in youngster and make them familiar with the miricals of science discovery.
The Genome editing Era (CRISPER Cas 9) : State of the Art and Perspectives fo...Anand Choudhary
Role of CRISPR/Cas9 in plant pathology
Production of disease resistance cultivars by editing the genome which is responsible for susceptibility factor for fungal and bacterial diseases.
By editing the genome which governs host pathogen interaction we can obtain incompatible interaction between host pathogen.
To improve the efficacy of bio control agents.
By editing the genome responsible for virus multiplication and virulence we can obtain virus free resistance cultivars.
The Genome-editing Era (CRISPER Cas 9) : State of the Art and Perspectives fo...ANAND CHOUDHARY
Role of CRISPR/Cas9 in plant pathology
Production of disease resistance cultivars by editing the genome which is responsible for susceptibility factor for fungal and bacterial diseases.
By editing the genome which governs host pathogen interaction we can obtain incompatible interaction between host pathogen.
To improve the efficacy of bio control agents.
By editing the genome responsible for virus multiplication and virulence we can obtain virus free resistance cultivars.
CRISPR is a new mechanism\tool to edit genes and in coming future it will provide us many new levels of success in curing of genetic disorders and modifying genes for human benifit
Introduction, History, components, cas9 protein structure and function,gRNA variants, Cas9 nuclease variants, CRISPR in bacteria as the immune system, mechanism, steps of working, Applications, and pros and cons.
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
1. CRISPR/CAS 9
A NEW TOOL FOR GENOME EDITING…
Dayalbagh
Educational Institute, Agra
PRESENTED BY
SHIVANI SINGH
M.SC FINAL SEM
2. CONTENT
INTRODUCTION
PIONEER SCIENISTS
HISTORY
BACKGROUND
DEFINATION
GENERAL IDEA BEHIND CRISPR
MECHANISM OF CRISPR/ CAS
USES OF CRISPR/CAS 9
3. INTRODUCTION
CRISPER stands for
Clustered regularly interspaced short
palindromic repeats.
What is Cas 9?
Cas9 (CRISPR associated protein 9, formerly called Cas5,
Csn1, or Csx12) is a 160 kD protein which plays a vital role
in the immunological defense of certain bacteria against
DNA viruses and plasmids.
4. 16 CRISPR Scientists & Pioneers: The Real Heroes
of Genome Engineering
Jennifer Doudna: The Mother of CRISPR
Emmanuelle Charpentier: CRISPR Co-Inventor
Feng Zhang: The Man Who Dared to Move Away from Bacteria
George Church: The Father of Genome Engineering
Francisco Mojica: Discovered the Existence of CRISPR Sequence in
Bacteria
Matthew Porteus: The Pioneer of Cell-Based CRISPR Therapies
David Liu: Introducing Base-Editor Enzymes
Stephen Tsang: Pioneer of Ophthalmological Genome Surgery Options
5. continued........
Kevin Esvelt: Changing the Game with Responsible Gene Drives
Prashant Mali: Taking the Multidisciplinary Approach
Stanley Qi: Developing the Next Stage of CRISPR Tech
Patrick Hsu: Expanding CRISPR Toolkit to RNA
Michel Sadelain: Novel Therapies Targeting Cancer
Andy Scharenberg: Editing T Cells
Jacob Corn: The Intersection of Genome Editing and DNA Repair
Alison Van Eenennaam: Using CRISPR to Edit Cattle
6. HISTORY
Where do CRISPR come from?
CRISPR was first discovered in archaea (and later in bacteria) by Francisco Mojica, a
scientist at the University of Alicante in Spain. He proposed that CRISPRs serve as part of
the bacterial immune system, defending against invading viruses. They consist of
repeating sequences of genetic code, interrupted by “spacer” sequences – remnants of
genetic code from past invaders. The system serves as a genetic memory that helps the
cell detect and destroy invaders (called “bacteriophage”) when they return. Mojica’s
theory was experimentally demonstrated in 2007 by a team of scientists led by Philippe
Horvath.
In January 2013, the Zhang lab published the first method to engineer CRISPR to edit
the genome in mouse and human cells.
7. BACKGROUND
How was it developed?
• Some bacteria have a similar, built-in, gene editing system to the
CRISPR-Cas9 system that they use to respond to invading
pathogens like viruses, much like an immune system.
• Using CRISPR the bacteria snip out parts of the virus DNA and
keep a bit of it behind to help them recognize and defend against
the virus next time it attacks.
• Scientists adapted this system so that it could be used in other
cells from animals, including mice and humans.
8. CRISPR-Cas9
Definition
CRISPR/Cas9 is a technique that
allows for the highly specific and
rapid modification of DNA in a
genome, the complete set of genetic
instructions in an organism.
9. GENERAL IDEA BEHIND CRISPR
CRISPR-Cas immunity is a natural process of bacteria and archaea.
CRISPR-Cas prevents bacteriophage infection, conjugation and
natural transformation by degrading foreign nucleic acids that
enter the cell.
CRISPR/Cas9 gene targeting requires a custom single guide RNA
(sgRNA) that contains a targeting sequence (crRNA sequence) and
a Cas9 nuclease-recruiting sequence (tracrRNA).
10. Specialized Cas proteins snip foreign DNA into small fragments
approximately 20 bp in length and paste them into contiguous stretches of
DNA known as CRISPR arrays. Separate Cas proteins then express and
process the CRISPR loci to generate CRISPR RNAs (crRNAs). Through
sequence homology, these crRNAs guide a Cas nuclease to the specified
exogenous genetic material, which must also contain a species-specific
sequence known as a protospacer adjacent motif (PAM). The CRISPR
complex binds to the foreign DNA and cleaves it to destroy the invader .
CONTINUED.....
12. CONTINUED.. BASIC TERMS RELATED TO
CRISPR CAS SYSTEM
are used in prokaryotic DNA editing involving CRISPR and
Cas9. These gRNAs are non coding short RNA sequences which bind to the complementary
target DNA sequences. Guide RNA first binds to the Cas9 enzyme and the gRNA sequence
guides the complex via pairing to a specific location on the DNA, where Cas9 performs its
endonuclease activity by cutting the target DNA strand.
is a region of non-coding DNA between genes. The terms intergenic spacer
(IGS) or non-transcribed spacer (NGS) are used particularly for the spacer DNA between the
many tandemly repeated copies of the ribosomal RNA genes. In bacteria, spacer DNA sequences
are only a few nucleotides long.
A protospacer adjacent motif (PAM) is a 2–6-base pair DNA sequence immediately following the
DNA sequence targeted by the Cas9 nuclease in the CRISPR bacterial adaptive immune system.
13. MOLECULAR MECHANISMS: ADAPTATION,
MATURATION AND INTERFERENCE
The CRISPR-Cas system acts in a sequence-specific manner by
recognizing and cleaving foreign DNA or RNA. The defence
mechanism can be divided into three stages:
(i) adaptation or spacer acquisition,
(ii) crRNA biogenesis, and
(iii) target interference
14. Acquisition of a new bacteriophage-derived spacer. Invading by a
bacteriophage for the first time, a new spacer derived from the
invading DNA and a repeat were incorporated into the CRISPR locus.
15. The immune mechanism of CRISPR-Cas system resisted invading DNA. When
the same bacteriophage invaded again, the CRISPR locus could transcribe
into a long pre-crRNA that was processed and generated small crRNAs. The
specific crRNA served as a guide for Cas proteins that recognized and
cleaved invading DNA.
16. Applications and Implications
CRISPR-Cas9 has a lot of potential as a tool for treating a range of
medical conditions that have a genetic component, including
cancer, hepatitis B or even high cholesterol.
Many of the proposed applications involve editing the genomes of
somatic (non-reproductive) cells but there has been a lot of
interest in and debate about the potential to edit germline
(reproductive) cells.
17. continued.....
Because any changes made in germline cells will be passed on
from generation to generation it has important ethical
implications.
The CRISPR/Cas9 is becoming a user-friendly tool for development
of non-transgenic genome edited crop plants to counteract
harmful effects from climate change and ensure future food
security of increasing population in tropical countries.
Potentials of CRISPR/Cas9 for improvement of crops cultivated in
tropical climates to gain resiliency against emerging pests and
abiotic stresses.
18. Applications of CRISPR CAS in
agriculture field...
Applications of
CRISPR/Cas9: CRISPR/Cas9
is currently used in genetic
engineering, agriculture,
and nutritional research.
19. Better targeting of CRISPR-Cas9
In most cases the guide RNA consists of a specific sequence of 20 bases.
These are complementary to the target sequence in the gene to be edited.
However, not all 20 bases need to match for the guide RNA to be able to
bind.
The problem with this is that a sequence with, for example, 19 of the 20
complementary bases may exist somewhere completely different in the
genome. This means there is potential for the guide RNA to bind there
instead of or as well as at the target sequence.
The Cas9 enzyme will then cut at the wrong site and end up introducing
a mutation in the wrong location. While this mutation may not matter at
all to the individual, it could affect a crucial gene or another important
part of the genome.
20. CONTINUE.....
Scientists are keen to find a way to ensure that the CRISPR-Cas9 binds
and cuts accurately. Two ways this may be achieved are through:
The design of better, more specific guide RNAs using our knowledge of
the DNA sequence of the genome and the 'off-target' behaviour of
different versions of the Cas9-gRNA complex.
The use of a Cas9 enzyme that will only cut a single strand of the target
DNA rather than the double strand. This means that two Cas9 enzymes
and two guide RNAs have to be in the same place for the cut to be
made. This reduces the probability of the cut being made in the wrong
place.
21. Different types of crispr cas system
1. CRISPR cas 9
Organisms: Streptococcus pyogenes, Streptococcus thermophilus, Staphylococcus aureus, Neisseria
meningitidis, Campylobacter jejuni
Size: ~1,000–1,600 aa
Guide spacer length: 18–24 nt
Total guide length: ~100 nt (sgRNA)
Cut: Blunt-ended dsDNA break
PAM: 3-NGG (SpCas9), 3-NNGRRT (SaCas9), 3-NNNNGATT (NmCas9)
The first and best-characterized single-protein CRISPR effector. Cas9 makes a blunt double-stranded
DNA break, which can then be repaired by either non-homologous end joining or homologous
recombination with a donor template DNA to create site-specific edits. Type II-A Cas9s generally have
high genome editing efficiency, but off-target cleavage at unintended genome sites can be a
disadvantage. Variants have been engineered to overcome these limitations, and type II-C Cas9s tend
to have naturally higher fidelity.
22. 2. CRISPR-Cas12
Organisms: Francisella novicida, Acidaminococcus sp., Lachnospiraceae sp., Prevotella sp.
Size: ~1,100–1,300 aa
Guide spacer length: 18–25 nt
Total guide length: 42–44 nt
PAM: 5-TTTN (FnCas12a)
Cut: 5 nt 5 overhang dsDNA break
Cas12 is a compact and efficient enzyme that creates staggered cuts in dsDNA. Cas12
processes its own guide RNAs, leading to increased multiplexing ability. Cas12 has also been
engineered as a platform for epigenome editing, and it was recently discovered that Cas12a
can indiscriminately chop up single-stranded DNA once activated by a target DNA molecule
matching its spacer sequence. This property makes Cas12a a powerful tool for detecting tiny
amounts of target DNA in a mixture.
23. 3. CRISPR-Cas13
Also known as C2c2 or CasRx
Organisms: Leptotrichia buccalis, Leptotrichia shahii, Ruminococcus flavefaciens, Bergeyella
zoohelcum, Prevotella buccae, Listeria seeligeri
Size: ~900—1,300 aa
Guide spacer length: 22–30 nt
Total guide length: 52–66 nt
Cut: ssRNA
PAM: 3-H (LshCas13a), 5-D and 3-NAN or NNA (BzCas13b), none (RfCas13d)
Cas13 is an outlier in the CRISPR world because it targets RNA, not DNA. Once it is activated by a
ssRNA sequence bearing complementarity to its crRNA spacer, it unleashes a nonspecific RNase
activity and destroys all nearby RNA regardless of their sequence. This property has been
harnessed in vitro for precision diagnostics. These systems can also be used for efficient,
multiplexable, and specific RNA knockdown or RNA sequence editing in mammalian cells. This
makes Cas13 a potentially significant therapeutic for influencing gene expression without altering
genome sequence.
24. The future of CRISPR-Cas9
Much research is still focusing on its use in animal models or isolated human
cells, with the aim to eventually use the technology to routinely treat diseases
in humans.
There is a lot of work focusing on eliminating ‘off-target’ effects, where the
CRISPR-Cas9 system cuts at a different gene to the one that was intended to be
edited.
The rapid progress in developing Cas9 into a set of tools for cell and molecular
biology research has been remarkable, likely due to the simplicity, high
efficiency and versatility of the system. Of the designer nuclease systems
currently available for precision genome engineering, the CRISPR/Cas system is
by far the most user friendly. It is now also clear that Cas9’s potential reaches
beyond DNA cleavage, and its usefulness for genome locus-specific recruitment
of proteins will likely only be limited by our imagination.