GSM8641423: CRISPR_T3; Homo sapiens; OTHER
ChIP-seq was performed as described previously (Schmidt D, Wilson MD, Spyrou C, Brown GD, Hadfield J, Odom DT.  ChIP-seq: using high-throughput sequencing to discover protein-DNA interactions.  Methods.2009;48:240-8)； Total RNA extracted using TRIzol was used for library preparation with an Illumina mRNA sequencing (mRNAseq) sample preparation kit.  Quality control of the RNA sequencing libraries was performed using a Bioanalyzer 2100 (Agilent, CA, USA) prior to paired-end sequencing on the HiSeq 6000 platform (Illumina, CA, USA);  ChIP-seq was performed according to the manufacturer's protocol (NEB, Shanghai, China). For CRISPR screening, Genomic DNA of day 0 cell samples and xenograft tumor samples were extracted with the Qiagen Blood & Cell Culture Midi Kit (Qiagen, Hilden, Germany). SgRNA cassettes were amplified from genomic DNA samples as the ﬁrst PCR, the amplicons resulting from the second PCR were extracted with beads (Beckman Coulter, USA), quantiﬁed, mixed, and sequenced using a NextSeq 500 instrument (Illumina, USA).