CRISPR-Cas9 editing of CHD8 is detrimental to B-ALL cells.

<p><b>(A)</b> Western blot showing loss of CHD8 expression or expression of a truncated protein following gene editing by CRISPR-Cas9 in B-ALL cells. Expression of truncated protein likely arises from an ORF in original reading frame following a new stop codon generated by gene editing. Chd8 sgRNA-1 and -2 cell lines were generated with sgChd8-2 construct, Chd8 sgRNA-4 was generated with sgChd8-3 construct, and Chd8 sgRNA-3 and -5 were generated with sgChd8-4 construct. <b>(B)</b> (Left) Bar graph showing results of <i>in vitro</i> growth competition assays with B-ALL cells following gene editing by CRISPR-Cas9. Constructs targeting Renilla luciferase (Ren) were used as negative controls. Ren-1 and -2 cell lines were generated using sgRen-2 construct, and Ren-3 was generated using sgRen-3 construct. Shown are averages ± SD. (Right) Detailed bar graph showing results of <i>in vitro</i> growth competition assays. <b>(C)</b> Kaplan-Meier curve showing survival of mice injected with B-ALL cells transduced with the indicated constructs. n = 10 per cohort. See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0143275#pone.0143275.s013" target="_blank">S6 Table</a> for statistics. ****P < 0.0001