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Gene knockout by CRISPR/Cas9

MT Ming-Feng Tsai
CP Charles B Phillips
MR Matthew Ranaghan
CT Chen-Wei Tsai
YW Yujiao Wu
CW Carole Williams
CM Christopher Miller
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Gene knockout by CRISPR/Cas9 was performed using the published protocol (Ran et al., 2013). In brief, the pSpCas9(BB) vector containing the 20-nucleotide guide sequence was transfected into HEK 293 cells. After two days of incubation, single cells were isolated by serial dilutions, and expanded for 2–4 weeks. Gene KO was assessed by sequencing and Western blot. Two sets of guide sequences (see Supplementary file 1) were used to rule out off-target effects.

Copyright and License information:  ©2016, Tsai et al
This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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