1. “CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system
Thank you.
Nucleeases will be used as a new tool that may revolutionize how we study genes.
Story of growing up on the farm.
Scientific discovory can be limited be the tools available.
Next few slides are a history of the DNA discoveries and tools used to manipulate DNA
Jason Kerkvliet, M.S.
Research Associate II
Hoppe lab
March 4, 2014

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

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

5. How does one use CRISPR? For NHEJ
CRISPR vector $65
4 (2 sgRNA sequences) oligos $40
BbsI $70
Total = $175

6. How does one use CRISPR? For HDR

7. CRISPR gRNA design tool
crispr.mit.edu

8. CRISPR/Cas9 can be used to edit more than one gene in parallel
Wang et al., Cell, 2013

9. Gene knock-down (siRNA) versus gene knock out
Shalem et al., Science 2014

10. Eliciting gene function using a CRISPR mediated genome screen
Shalem et al., Science 2014