The Biggest Game Changer In Biology Since PCR .

1. Name : BASEL , Mahmoud , Ahmed , Roaa
University : Kafr El-Sheikh , Egypt
Study in : Faculty of Fisheries And Aquaculture Sciences
Department : Fish Processing And Biotechnology
“ And mankind have not been given of
knowledge except a little.” Israa 85
«‫ا‬َ‫م‬ َ‫و‬َ‫ن‬ِ‫م‬ ْ‫م‬ُ‫ت‬‫ي‬ِ‫ت‬‫و‬ُ‫أ‬ِ‫م‬ْ‫ل‬ِ‫ع‬ْ‫ال‬َ‫ّل‬ِ‫إ‬
ً‫يل‬ِ‫ل‬َ‫ق‬»‫اّلسراء‬ ‫سورة‬85

2. The Biggest Game Changer
In Biology
Since PCR

3.  What Is CRISPR Mean?
• “ CRISPR ” Stands For :
Clustered
Regularly
Interspaced
Short
Palindromic
Repeats
• It has rapidly become one of the most popular
approaches for genome engineering .

4. The Tri Top Methods of genome Editing
TALEN
CRISPR/cas9
ZFN

5. HISTORY
 1987 ..
 2002 ..
 2007 ..
 2011 ..
 2012 ..
 2013 ..
Agriculture Application of CRISPR raised 11 $ million
CRISPR Sequences were First Discovered In E.coli ( Ishino )
Identification Of cas gene associated with DNA repeats in
Prokaryotes ( Jansen )
CRISPR Provides acquired resistance against Viruses in
Prokaryotes ( Barrangou )
Idea of using CRISPR cas9 as genome Engineering tool was
published by Jennifer Doudna and Emanuelle Charpenter
CRISPR is used in mouse and Human cells by Feng Zhang

6. What makes CRISPR system the ideal genome
engineering technology :
1) High Potency and Specificity .
2) Broad applicability to both in vivo and ex vivo
applications .
3) Simple editing tools ( guide RNA + Protein ).
4) Ability to address any site in the genome or
foreign genome .
5) Malfunctional programmability :
Delete , Insert , Repair genes and Muliple DNA sites .

7.  Conflict For CRISPR
• Jennifer Doudna and Emmanuelle Charpentier re-engineered
the Cas9 endonuclease into a more manageable two-component
system by fusing the two RNA molecules into a "single-guide
RNA" that, when combined with Cas9, could find and cut the
DNA target specified by the guide RNA. By manipulating the
nucleotide sequence of the guide RNA, the artificial Cas9
system could be programmed to target any DNA sequence for
cleavage.
• Feng Zhang's and George Church's groups simultaneously
described genome editing in human cell cultures using CRISPR-
Cas9 for the first time. It has since been used in a wide range of
organisms, including baker's yeast , zebra fish , fruit flies ,
nematodes , plants , mice , monkeys and human embryos.

8. CRISPR pioneers (from left to right):
George Church , Jennifer Doudna , Feng Zhang and Emmanuelle Charpentier
Were Ended With Drawn

9.  DISCOVERY
• “ CRISPRs “ were first discovered in Archaea and later in
Bacteria by Francisco Mojica who proposed that CRISPRs
serve as part of the Bacterial immune system , defending
against invading viruses .
• The system serves as a Genetic Memory that helps the cell
detect and destroy when they return .
• In Jan 2013 , Feng Zhang at the Broad Institute And MIT
published the first method to engineer CRISPR to edit the
genome in mouse and human cells .

10. Action Of CRISPR In Bacteria
 The CRISPR immune system works to protect Bacteria
from repeated viral attack via 3 basic steps :
1) Adabtation
2) Production of crRNA
3) Targeting

11. Different CRISPR-Cas system in Bacterial Adaptive Immunity
Class 1 (CRISPR-Cas 3 & CRISPR-Cas 10)
 uses several Cas proteins and the crRNA .
Class 2 ( CRISPR-Cas 9 & CRISPR-Cpf 1 )
 Employ a large single component cas 9 protein in
conjunction with crRNA and tracRNA .

12. Components Of CRISPR
• Protospacer Adjacent Motif ( PAM )
• CRISPR-RNA (crRNA )
• Trans-Activating crRNA ( tracrRNA )

13.  CRISPR-Cas 9
• CRISPR associated “ Cas “ proteins .
• “ Cas 9 “ is the Nucleus guided by the trans-activating
crRNA to cleave specific DNA sequences .
• It is the Simplest , Most Versatile and Precise method of
genetic Manipulation .

14.  MECHANISM
• “ CRISPR “ scans the genome looking for the right location and then uses the
Cas9 protein as molecular scissors to snip through the DNA .
• “ Cas9 endonuclease “ guide RNAs to direct it to a particular sequence to be
edited. The genetic sequence of the RNA matches the target sequence of the
DNA that has to be edited.
• When Cas9 cuts the target sequence, the cell repairs the damage by replacing
the original sequence with an altered version.

15.  What are the pros of Gene editing?
• “ CRISPR “could be used to modify disease-causing genes
in embryos brought to term, removing the faulty script
from the genetic code of that person’s future descendants
as well.
• Genome editing could potentially decrease, or even
eliminate , the incidence of many serious genetic diseases,
reducing human suffering worldwide.
• It might be possible to install genes that
offer lifelong protection against infection .

16.  What are the cons of Gene editing?
• Risky human experimentation .
• Altering one gene could have unforeseen and wide spread effects on other
parts of the genome .
• It will become a tool for selecting desired characteristics
such as intelligence and attractiveness .
• Many consider genome alterations to be unethical , advocating that we
should let nature run its course.

17.  8 Ways CRISPR-Cas9 Can Change the World
1. Remove malaria from mosquitos.
2. Treating Alzheimer’s disease.
3. Treating HIV. The HIV virus inserts its DNA into the cells of the
human host.
4. Develop new drugs.
5. Livestock. CRISPR/Cas9 has been utilized in China to delete genes.
6. Agricultural crops . to improve crop disease resistance and
environmental stress tolerance in plants.
7. Develop new cancer treatments -modify immune cells.
8. Reduce our need for plastic. CRISPR can be used to manipulate a
type of yeast that transforms sugars into hydrocarbons, which can be
used to make plastic .

18. Companies

19. Software and Databases

20. The End of The Presentation
but Not The End Of CRISPR