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“CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system Thank you. Nucleeases will be used as a new tool that.

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Presentation on theme: "“CRISPR genome editing” Precise gene regulation/modification using the simple CRISPR/Cas9 system Thank you. Nucleeases will be used as a new tool that."— Presentation transcript:

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

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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

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