Krishnananda Pralhad Ingle (PhD Agricultural BIotechnology) Dr. PDKV, Akola

1. CRISPR/Cas9 GENOME EDITING
Presented by
Krishnananda Pralhad Ingle
(PhD Agricultural Biotechnology)
Biotechnology Centre, Dr. Panjabrao Deshmukh
Agricultural University, Akola (MS) India 444104

2. Genome Editing Tools
Genome editing is the process of precisely modifying the nucleotide sequence of the genome
CRISPR-Cas9 is a unique technology that enables geneticists & medical researchers to edit parts of
the genome by removing, adding or altering sections of DNA sequence
Protein Dependent DNA
CleavageCleavage
ZFN TALEN
RNA Dependent DNA
Cleavage (CRISPR)
Oligo-nucleotide-directed
mutagenesis (ODM)
Cleaved Genomic Site
Repair

3. Timeline outline
CRISPR elements and associated
genes identified
CRISPR proposed to be a bacterial
adaptive immune system
CRISPR/Cas 9 used to edit the
targeted genes in both humans
and mouse cells
USDA determine that CRISPR/Cas9
edited crops will not be regulated
as GMO
CRISPR repeat first observed
in bacterial repeats
20021987 2005 2006 2007 2010 2013 2015 2016 2017
CRISPR spacer identified as
foreign DNA
Discovery that CRISPR/Cas
imparts resistance to
bacterial phages
CRISPR/Cas identified as
bacterial and archael
immune system
CRISPR/Cas9 used to develop
virus resistance tomato plants
US patent office awarded
CRISPR/Cas9 patent to Broad
Institute

4. Main Actors in Patent War

5. CRISPR System
The part of the Bacterial immune system which detects and recognize the foreign
DNA and cleaves it.
1. TheCRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)
2. Cas (CRISPR- associated) proteins can target and cleave invading DNA
in a sequence – specific manner.
A CRISPR array is composed of a series of repeats interspaced by spacer
sequences acquired from invading genomes.

6. CRISPR Components
Protospacer adjacent
Motif (PAM)
CRISPR-RNA (crRNA)
trans-activating crRNA
(tracrRNA)

7. Steps involved in CRISPR/Cas9

8. Schematic representation of Cas9/sgRNA system

9. Evolutionary classification of CRISPR–Cas systems

10. Model of CRISPR/Cas System

11. Molecular mechanism for driving gene editing in CRI
SPR-Cas9 technology

12. Potential Usage
Microbial Research
Cell line
development
Animal Research
Crop Improvement
Functional Genomics
Drug Designing
Human Gene
Therapy
CRISPR/Cas9
Genome Engineering

13. 1. Non-target effects- the possibility of non target effects of
synthetic nucleases during genome editing.
2. Regulation of plants created by genome editing – not a transgenic
Valuation of genome editing systems

14. Limitations of CRISPER- Cas9 System
1. How to deliver gene editing to the right cells
2. Its offsite activity as a little data available regarding
offsite activity in plants.
3. Plant Viral vectors can’t be applied as genome editing3. Plant Viral vectors can’t be applied as genome editing
tool
4. Spread of edited strains in the ecosystem can cause
serious problem to ecosystem.

15. Conclusion
CRISPR/Cas9 system which offers several avenues for
genome editing in diverse species and opportunity to
make the best use of it to improve the process and
product in their field of research for the betterment ofproduct in their field of research for the betterment of
humankind, quality food supply and food security to
increasing population.
CRISPER- Cas ………. ???????

16. Thank You…!