Successfully reported this slideshow.
We use your LinkedIn profile and activity data to personalize ads and to show you more relevant ads. You can change your ad preferences anytime.
CRISPR-CAS 9
Hemant Jain and Bhakt singh
keneddy cherukula .. … . , ..
What is CRISPR
1. It is a bacterial defence mechanism
2. First discovered in ecoil in 1980s
Cluster regularly interspaced short
palindrome repeat
Clustered regularly interspaced short
palindromic repeats
• Spacer dna – ???
• It was found in 2000 that spacer dna matches perfectly with
bacteriophage dna
• Also the genes associ...
CAS genes produces cas protien complex
• This CAS complex has 2 parts
1. hilecase part
2. nucleases part
Therefore it acte...
In case when viral dna enters the host cell
• There will be 3 scenarios
Scenario 1
Scenario 2
The CRISPR gene
would transcript and
translate Toform CAS
protien complex
That denaturated
the viral DNA by
ide...
Scenario 3
• If the viral DNA dose not
Matches with the
interspaced DNA then a
different class of CAS
protien complex woul...
EUREKA
• Realizing this fact the scientist could now use it to activate or
incorporate new genes in the host
• Scientist Jennifer
doudna and
Emmanuelle
Charpentier
identified CAS 9 in
their labs while
working on
steptococcus
pyogen...
CAS 9
• It has crRNA
(SPACER
DNA ) and
trRNA that
holds crRNA
in place
• The scientists
modifiedthis system
by using cas 9
protien and use the
host cell’s RNA and
by connecting it to
tracrRNA i...
• Due to this
the CAS 9
detects the
required DNA
sequence by
using the
host’s own
RNA as a
template and
cuts it out
• Afte...
The RNA in CAS 9 recognises the DNA
sequence and cleaves it
After cleaving the DNA reparation/ ligation
takes place
Hence a knockout could be produced
By this system a new gene could be induced
at the place of deleted gene
Advantages of CRISPR/CAS over ZNF and
TALEN mutagenesis stratagies
• 1. Target design simplicity. Because the target speci...
• 3. Multiplexed mutations. Mutations can be introduced in multiple
genes at the same time by injecting them with multiple...
Disadvantages of CRISPR-CAS 9
• 1. Off-site effects. Mutation introduced at non-specific loci with similar, but not
identi...
CRISPR has
Many applications
CRISPR in medicine HIV/AIDS
Other medical applications
Muscular Dystrophy
Cancer
And Many more..
CRISPR-Cas9 in functional genomic screening
• Functional genomic screening is largely used for identifying the
essential g...
CRISPR-Cas9 in correction of genetic disorders
• One of the most exciting applications of the CRISPR-Cas9 is the
possibili...
CRISPR-Cas9 in the treatment of infectious diseases
• Considering that the CRISPR-Cas system originally functions as an
an...
CRISPR-Cas9 in the generation of animal models
• gene targeting based on homologous recombination and embryonic
stem cells...
Crispr
Crispr
Crispr
Crispr
Upcoming SlideShare
Loading in …5
×

Crispr

2,814 views

Published on

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

Published in: Science
  • Who Else Wants To Permanently Cure Their Uterine Fibroids and Achieve LASTING Freedom From PCOS Related Symptoms? learn more... ★★★ http://t.cn/Aig7V1M7
       Reply 
    Are you sure you want to  Yes  No
    Your message goes here

Crispr

  1. 1. CRISPR-CAS 9 Hemant Jain and Bhakt singh keneddy cherukula .. … . , ..
  2. 2. What is CRISPR 1. It is a bacterial defence mechanism 2. First discovered in ecoil in 1980s
  3. 3. Cluster regularly interspaced short palindrome repeat
  4. 4. Clustered regularly interspaced short palindromic repeats
  5. 5. • Spacer dna – ??? • It was found in 2000 that spacer dna matches perfectly with bacteriophage dna • Also the genes associated with crispr gene was discovered i.e CAS genes
  6. 6. CAS genes produces cas protien complex • This CAS complex has 2 parts 1. hilecase part 2. nucleases part Therefore it acted As immune system Of bacteria.
  7. 7. In case when viral dna enters the host cell • There will be 3 scenarios Scenario 1
  8. 8. Scenario 2 The CRISPR gene would transcript and translate Toform CAS protien complex That denaturated the viral DNA by identifying by the copy of host RNA as a template for recognition
  9. 9. Scenario 3 • If the viral DNA dose not Matches with the interspaced DNA then a different class of CAS protien complex would be produced that will break the viral DNA and incorporate it as an interspaced DNA among the palindromes . • Therefore these interspaced DNA acts as the history of old infection and hence developes further immunity .
  10. 10. EUREKA • Realizing this fact the scientist could now use it to activate or incorporate new genes in the host
  11. 11. • Scientist Jennifer doudna and Emmanuelle Charpentier identified CAS 9 in their labs while working on steptococcus pyogenes
  12. 12. CAS 9 • It has crRNA (SPACER DNA ) and trRNA that holds crRNA in place
  13. 13. • The scientists modifiedthis system by using cas 9 protien and use the host cell’s RNA and by connecting it to tracrRNA i.e. trcrRNA-crRNA chimera also called grRNA.
  14. 14. • Due to this the CAS 9 detects the required DNA sequence by using the host’s own RNA as a template and cuts it out • After that DNA reparation takes place
  15. 15. The RNA in CAS 9 recognises the DNA sequence and cleaves it
  16. 16. After cleaving the DNA reparation/ ligation takes place
  17. 17. Hence a knockout could be produced
  18. 18. By this system a new gene could be induced at the place of deleted gene
  19. 19. Advantages of CRISPR/CAS over ZNF and TALEN mutagenesis stratagies • 1. Target design simplicity. Because the target specificity relies on ribonucleotide complex formation and not protein/DNA recognition, gRNAs can be designed readily and cheaply to target nearly any sequence in the genome specifically. • 2. Efficiency. The system is super-efficient. Modifications can be introduced by directly injecting RNAs encoding the Cas protein and gRNA into developing mouse embryos. This eliminates the long and laborious processes of transfecting and selecting mouse ES cells that are required to create targeted mutant mice using classical homologous recombination techniques.
  20. 20. • 3. Multiplexed mutations. Mutations can be introduced in multiple genes at the same time by injecting them with multiple gRNAs
  21. 21. Disadvantages of CRISPR-CAS 9 • 1. Off-site effects. Mutation introduced at non-specific loci with similar, but not identical, homology to the target sites are one of the most important complication of these technologies. These can be difficult to identify and require scanning the genome for mutations at sites with sequence similarity to the gRNA target sequence. • 2. Mosaicism. Mice with a mutant allele in only some of their cells can be produced , because the nucleases may not necessarily cut the DNA at the one cell stage of embryonic development. • 3. Multiple alleles. Healing of the nuclease cleavage site by non-homologous end joining can produce cohorts of mice with different mutations from the same targeting constructs, requiring genome sequencing to verify the nature and position of the specific mutation. The production of mice with mosaics of multiple mutations, also, is possible, and breeding may be required to segregate and isolate mice that carry single mutations. The production of mice with multiple variants also creates phenotyping bottlenecks.
  22. 22. CRISPR has Many applications
  23. 23. CRISPR in medicine HIV/AIDS
  24. 24. Other medical applications Muscular Dystrophy Cancer And Many more..
  25. 25. CRISPR-Cas9 in functional genomic screening • Functional genomic screening is largely used for identifying the essential genes for a specific cellular process.
  26. 26. CRISPR-Cas9 in correction of genetic disorders • One of the most exciting applications of the CRISPR-Cas9 is the possibility of curing genetic diseases. • Eg . Could event cataract by inducing mutation in CYRC gene • Similarly could also cure cystic fibrosis by targeting multiple genes
  27. 27. CRISPR-Cas9 in the treatment of infectious diseases • Considering that the CRISPR-Cas system originally functions as an antiviral adaptive immune system in bacteria, this system could be used for treating infectious diseases by eradicating pathogen genomes from infected individuals.
  28. 28. CRISPR-Cas9 in the generation of animal models • gene targeting based on homologous recombination and embryonic stem cells has been used as the typical approach for animal genome modification, which has played indispensable roles in making a causal link between genomic mutations and phenotypes during development and in disease. However, gene targeting has limited applications in some organisms due to time-consuming procedures and the lack of available embryonic stem cells

×