“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 IIHoppe lab
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system
DNA repair mechanisms: gene editing efficiency is enhanced by creating breaks in DNA
How does one use CRISPR? For NHEJ
• CRISPR vector $65• 4 (2 sgRNA sequences) oligos $40• BbsI $70Total = $175
How does one use CRISPR? For HDR
crispr.mit.edu CRISPR gRNA design tool
CRISPR/Cas9 can be used to edit more than one gene in parallel
Wang et al., Cell, 2013
Shalem et al., Science 2014
Gene knock-down (siRNA) versus gene knock out
Eliciting gene function using a CRISPR mediated genome screen
Shalem et al., Science 2014
http://www.addgene.org/CRISPR/
http://www.addgene.org/CRISPR/plant/
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