Genome Editing BY CRISPER

1. Submitted By:
Osama Alam
Department of Bio-Technology
University of Science & Technology, Bannu
September 2022

2. Genome Editing
Presented by:
Osama Alam
Presented to:
Sir Sami Sb

3. CRISPR-CAS 9 Method
• CRISPR Associated Nuclease
•Definition
It is is a unique method of genome editing that helps to find a specific bit
of DNA inside a cell.
It edits genes by highly precisely cutting DNA and then letting natural
DNA repair processes to take over.
• Clustered
• Regulory
• Interspaced
• Short
• Palindrome Repeats

4. CRISPR
It is a short segment of DNA containing short repetitions of base
sequences, involved in the defence mechanisms of prokaryotic
organisms to viruses.
It is found in the genomes of prokaryotic organisms such as bacteria and
archaea.
It helps to detect and destroy DNA from similar bacteriophages

5. Mechanism
oWhen bacteriophage attack on bacteria the first stage of the
immune response is to capture phage DNA and insert it into a
CRISPR locus in the form of a spacer. Cas1 and Cas2 .
oSpacer DNA +Palindrome =CRISPR Locus Array
osDNA is a region of non coding and they are mostly transposan
oWhen viral DNA is inserted in bacteria genome it is transcribe.

8. Components
CAS-9 CRISPER Associated Enzyme:
• It helps to precise cut in DNA double strand and shift the
target gene.
Guided RNA:
• It helps to guide the RNA for precise cut.
• They contain such sequences that are complement to DNA
sequence.
• It cut in such places that we need of there edition.
• Trans Activating CRISPER
• It helps to Guide the Guided RNA sequence.

10. Designing sgRNA for CRISPR
Experiments

11. PAM sequences
• Each Cas nuclease binds to its target sequence only in presence of a
specific sequence, called protospacer adjacent motif (PAM).
• The nuclease cuts 3-4 nucleotides upstream of the PAM sequence

12. How CRISPER Work
i. RNA can be designed for many unique sequences of
DNA.
ii. Targeted DNA may be faulty.
iii. Could be theraptic.

14. Application