CRISPR-Cas9 Cas9

Genome Target sequence

Editing
Target loci

Md. Qamrul Islam

Registration No. 2015431023
Dept. of Genetic Engineering and Biotechnology
Shahjalal University of Science and Technology, Sylhet
 Genome Editing
• DNA is inserted

• Deleted

• Replaced

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 Advancements in Genome Editing
1989 1998 2009
HR mediated ZFNs TALENs
targeting

1992 2000 2013

Cre-lox Bacterial CRISPR/Cas
CRISPR
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 What is CRISPR ?
• Clustered regularly interspaced short palindromic
repeats

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 Discovery of CRISPR
• Osaka University researcher Yoshizumi Ishino in
1987.

ENEMIES

FIGHTING FOR
EXISTANCE
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 Components of CRISPR Locus
A CRISPR Locus
Spacers

Cas genes Leader Repeat

A CRISPR Array

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Components of CRISPR-Cas9

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Natural Microbial CRISPR System

Phage infection

Spacer Acquisition
4 Steps
crRNA Biogenesis and
Processing

Target Degradation

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 Natural Microbial CRISPR System

1. Phage infection 2. Spacer Acquisition

Hsu et al. 2014

Figure: Mechanism of CRISPR mediated immunity in bacteria
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 Natural Microbial CRISPR System

3. crRNA Biogenesis and Processing

Hsu et al. 2014

Figure: Mechanism of CRISPR mediated immunity in bacteria
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 Natural Microbial CRISPR System

4. Target Degradation

Hsu et al. 2014

Figure: Mechanism of CRISPR mediated immunity in bacteria
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Application in Genome Engineering

Figure: DNA repair after double-strand break

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Application in Knockdown/Activation

Figure: Cas9 system for gene activation and repression

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 Strategies of using CRISPR/Cas

Figure: A general scheme using CRISPR/Cas in genome editing

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Latest Developments in CRISPR Research

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 Future Prospects
Ecological Gene therapy Screen for drug
vector control target ID

CRISPR-Cas9 mediated genome engineering

Pathogen gene
Pathway engineering Crops, animals
disruption

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 Conclusion
• Medical applications

• Crop editing

• Gene drives

• Synthetic biology

• Further investigation

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Thank you

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