CRISPR screen identifies genes that sensitize AML cells to double negative T cell therapy [RNA-Seq]

Here, we used a targeted CRISPR/Cas9 screen to identify genes that confer susceptibility of AML cells to DNT cell therapy. Total RNA was extracted from AML3 cells using the RNeasy Plus Mini Kit (Qiagen Cat. 74314) following manufacture’s guidelines. On column DNase I digestion was performed for 15 mins at room temperature and RNA purity was assessed by Nanodrop (260/280, > 2.0). RNA-seq libraries were prepared using the SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian (634418, Takara). Libraries were verified using TapeStation (Tape 2200, Agilent Technologies) and 150-bp paired-end sequencing was performed