Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution
Title:
Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution
Author:
Lee, Joonsun
Personal Author:
Publication Information:
Cambridge, MA MyJoVE Corp 2016
Physical Description:
online resource (697 seconds)
Series:
Genetics
General Note:
Title from resource description page
Abstract:
Here, we present a protocol to optimize CRISPR-Cas9 to achieve a higher specificity without the loss of on-target activity. We use a directed evolution approach called Sniper-screen to find a mutant Cas9 with the desired characteristics. Sniper-Cas9 is compatible with truncated single-guide RNAs and delivery in a ribonucleoprotein format, well-known strategies for achieving higher specificities.
Reading Level:
For undergraduate, graduate, and professional students
Subject Term:
Electronic Access:
https://www.jove.com/t/59202