title: HEK293T_A8e
design: HEK293T cells were seeded into the 6-well plate at an appropriate density. The negative control group was transfected with 6 g guide RNA plasmids and kept in thick tinfoil all the time. The blue light was turned 24h later, and cells were cultured for another 48h. Cells were washed in pre-cooled DPBS and collected in a 15 mL tube. After centrifugation in 1200 r.p.m. for 10 min, cells were resuspended in TRIzol and frozen in liquid nitrogen.Total RNA was extracted from the cells according to the manufacturer's instructions of TRIzol (Invitrogen), and genomic DNA was removed using DNase I (TaKara). RNA degradation and contamination were monitored on 1% agarose gels. The RNA library was sequenced using Illumina NovaSeq 6000 sequencer in 2150bp read length. The raw paired-end reads were trimmed and quality controlled by fastp with default parameters. Then clean reads were separately aligned to the reference genome with orientation mode using HISAT2 software. StringTie assembled the mapped reads of each sample in a reference-based approach. SNP(Single Nucleotide Polymorphisms) calling analysis was performed by Sentieon Genomics Suite.