“crispr genome editing” precise gene regulation/modification using the simple crispr/cas9 system

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“CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system March 4, 2014 Jason Kerkvliet, M.S. Research Associate II Hoppe lab

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“CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system . Jason Kerkvliet , M.S. Research Associate II Hoppe lab. March 4 , 2014. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system. - PowerPoint PPT Presentation

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Page 1: “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system

“CRISPR genome editing”Precise gene regulation/modification using

the simple CRISPR/Cas9 system

March 4, 2014

Jason Kerkvliet, M.S.Research Associate IIHoppe lab

Page 2: “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system

Page 3: “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system

DNA repair mechanisms: gene editing efficiency is enhanced by creating breaks in DNA

Page 4: “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system
Page 5: “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system

How does one use CRISPR? For NHEJ

• CRISPR vector $65• 4 (2 sgRNA sequences) oligos $40• BbsI $70Total = $175

Page 6: “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system

How does one use CRISPR? For HDR

Page 7: “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system

crispr.mit.edu CRISPR gRNA design tool

Page 8: “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system

CRISPR/Cas9 can be used to edit more than one gene in parallel

Wang et al., Cell, 2013

Page 9: “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system

Shalem et al., Science 2014

Gene knock-down (siRNA) versus gene knock out

Page 10: “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system

Eliciting gene function using a CRISPR mediated genome screen

Shalem et al., Science 2014

Page 11: “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system

http://www.addgene.org/CRISPR/

Page 12: “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system

http://www.addgene.org/CRISPR/plant/