RNA-seq analysis of knockout cells
Oligo-dT primers from APExBIO (China) were employed for mRNA transversion to acquire cDNA. Second cDNA chain synthesis was achieved by amplifying the obtained cDNA. Purify cDNA products by AMPure XP system (Beckman Coulter). PCR amplification was performed to enrich library fragments, which were then selected based on a size range of 350550 bp. The quality of the library was assessed using an Agilent 2100 Bioanalyzer (Agilent). Subsequently, the library was sequenced using the Illumina NovaSeq 6000 platform, generating raw reads.