"crispr" gene editing tool

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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 : Dr. Hari Pathak

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Page 1: "CRISPR" Gene Editing Tool

BIOTECHNOLOGY DEPARTMENTSEMINAR TOPIC: “CRISPR” GENE

EDITING TOOL (New Era in Molecular Biology)

BY, ZAINAB SHAHIDFINAL YEAR1306454040DATE:21.MARCH.2017 SUBMITTED TO :

Dr. Hari Pathak

Page 2: "CRISPR" Gene Editing Tool

INTRODUCTION..

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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.

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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.

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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.

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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.

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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.

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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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