The document summarizes a seminar presentation on CRISPR gene editing. It discusses how CRISPR/Cas systems in bacteria provide adaptive immunity by integrating fragments of viral DNA. It then describes some applications of CRISPR technology, including inactivating genes in human and other cells, modifying crops and mosquitoes. Recent developments include licensing of CRISPR kits and using CRISPR/Cas9 to introduce targeted mutations in model organisms and cell lines. Chinese researchers also used CRISPR/Cas9n to produce cattle with increased resistance to bovine tuberculosis by inserting a new gene.

1. BIOTECHNOLOGY DEPARTMENT
SEMINAR TOPIC: “CRISPR” GENE
EDITING TOOL (New Era in Molecular
Biology)
BY, ZAINAB SHAHID
FINAL YEAR
1306454040
DATE:21.MARCH.2017
SUBMITTED TO :

2. INTRODUCTION..

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8. The CRISPR/Cas system
Their function was confirmed in 2007 by Barrangou and
colleagues, who demonstrated that S. thermophilus
can acquire resistance against a bacteriophage by
integrating a genome fragment of an infectious
virus into its CRISPR locus.
In 2005, three independent research groups showed
that some CRISPR spacers are derived from phage
DNA and extra chromosomal DNA such as plasmids.

9. The CRISPR/Cas system
• In effect, the spacers are fragments of DNA
gathered from viruses that previously tried to
attack the cell.
• The source of the spacers was a sign that the
CRISPR/cas system could have a role in adaptive
immunity in bacteria.

10. Applications
• By the end of 2014 some 600 research papers
had been published that mentioned CRISPR.
• The technology had been used to functionally
inactivate genes in human cell lines and cells,
• To study Candida albicans
• To modify yeasts used to make biofuel and to
genetically modify crop strains.
• CRISPR can also be used to change mosquito so
they cannot transmit diseases such as malaria.

11. Recent
• As of November 2013, SAGE Labs had
exclusive rights from one of those companies
to produce and sell genetically engineered rats
and non-exclusive rights for mouse and rabbit
models.
• By 2015, Thermo Fisher Scientific had
licensed intellectual property from ToolGen to
develop CRISPR reagent kits.

12. Applications as a Genome-editing and
Genome Targeting Tool
• In 2012 , the CRISPR/Cas9 system has been widely adopted.
• This has already been successfully used to target important genes in many cell
lines and organisms, including human , bacteria , zebrafish , C. elegans, plants ,
Xenopus tropicalis, yeast , Drosophila , monkeys , rabbits , pigs , rats and mice .
• Several groups have now taken advantage of this method to introduce single point
mutations (deletions or insertions) in a particular target gene, via a single gRNA
• The future of CRISPR/Cas9: due to the simplicity, high efficiency and versatility of
the system.
• Cas9’s potential reaches beyond DNA cleavage, and its usefulness for genome
locus-specific recruitment of proteins is possible.

13. Chinese researchers develop TB resistant
cow
• Scientists have used gene-editing technology CRISPR/Cas9
for the first time to successfully produce live cows with
increased resistance to bovine tuberculosis
• used a modified version of the CRISPR system called CRISPR/Cas9n to
insert a new TB resistance gene-NRAMP1 into the genome of bovine foetal
fibroblasts which is a cell derived from female dairy cows. These cells were
used as donor cells in a process called SCNT, where the nucleus of a donor
cell carrying the new gene is inserted into an egg cell.
Results revealed that NRAMP1 had successfully integrated into the
genetic code at the targeted region in all of the calves. When exposed to
Mycobacterium bovis – (the bacterium that causes bovine TB)- the
transgenic animals showed an increased resistance to the bacteria
measured by standard markers of infection in a blood sample.

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