Figure 2.

The expression of XIST and X-linked genes after the knockout of D-repeat. (A) The sgRNA- mediated D-repeat mutation were analyzed by PCR and agarose gel. Genomic DNA was extracted from sgRNA transfected and control cells, subjected to PCR and 2% agarose gel analysis. M, marker III. KO, knockout of D-repeat. Con, empty vector. (B) The mutation efficient of were analyzed by T-vector sequences analysis. WT represent the sequence of 7.6 tandem repeat units in D-repeat. The PAM sequence is marked in purple and the off-target sites marked in green. The sequence of region deletions is under gray background and the length of deletions is noted to the right of each sequence (- deletion). The sgRNA targeted TRs were demonstrated in red underlined.(C) The expression of XIST and X-linked genes were investigated by q-PCR. Data are shown as mean ±SEM (n = 3). Asterisks denote significant differences between Con and KO samples; *p < 0.05,**p < 0.001.