“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. 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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Precise Genome Editing In Mammalian Cells using the simple and Versatile CRISPR/Cas9 system: The Beginning of a New Era in Biological Sciences

CRISPR genome editingPrecise gene regulation/modification using the simple CRISPR/Cas9 system March 4, 2014

Jason Kerkvliet, M.S.Research Associate IIHoppe lab

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 1Clustered 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 $654 (2 sgRNA sequences) oligos $40BbsI $70Total = $175

How does one use CRISPR? For HDR

crispr.mit.eduCRISPR 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 2014http://www.addgene.org/CRISPR/

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