Supplementary Figure 7: dCas9–KRAB–MeCP2-mediated repression is highly specific in human cells.

(a) HEK293T cells were transfected with a gRNA targeting the CXCR4 gene along with the indicated dCas9 repressor. Expression of the target gene CXCR4 and several nearest neighboring genes, including MCM6, DARS, THSD7B, HNMT, and SPOPL, were examined. These genes are located at approximately -274, -208, 651, 1849, and 2387 kb, respectively, away from CXCR4. (b) Expression of the target gene SYVN1 and several nearest neighboring genes, including ZNHIT2, FAU, MRPL49, SPDYC, and CAPN1, were examined. These genes are located at approximately -10, -6, -5, 42, and 53 kb, respectively, away from SYVN1. For a-b, data are shown as mean ± s.e.m. n = 4 biologically independent samples (cell cultures). One-sided Student T-test was used to perform statistical comparison. * p < 0.05 v.s. dCas9-KRAB. (c) RNA-seq analysis of HEK293T cells transfected with gRNA targeting CXCR4 along with dCas9, dCas9-KRAB or dCas9-KRAB-MeCP2. Shown are density plots indicating the fold changes in gene expression from different groups. CXCR4 expression is indicated with blue, orange, or red dot and control is indicated with a black dot. n = 2 biologically independent samples (cell cultures).