Tips for CRISPR Gene Editing in Mice
Type
Blog Post
Published
June 28, 2016, 10:59 a.m.
...screening for the best gRNA
Figure 1: Validation of double strand break (DSB) mediated homology dependent...as control (Cetn1/sgRNA1), 2; darker than control, 1; very dark). Brighter fluorescence indicates a more...and greatest in scientific content.
References
1. Cho, Seung Woo, et al. "Analysis of off-target effects...repair (HR, homologous recombination; SSA, single strand annealing) took place and reconstituted the EGFP...site. (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences...efficacy of your gRNAs is then a simple matter of transfecting HEK293T cells with your modified pCAG-EGXXFP...fluorescence, the better that gRNA is at causing a double stranded break at the target site, allowing the two halves...