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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
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 $70
Total = $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
Gene knock-down (siRNA) versus gene knock out
Shalem et al., Science 2014
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/