RNA-seq of Jurkat cells treated with RENDER-CRISPRoff targeting CD55 replicate 1
CLTA-GFP HEK293T cells were maintained for 24 days post-treatment of RENDER-CRISPRoff v3 targeting CLTA. Jurkat cells were maintained for 24 days post-treatment of RENDER CRISPRoff v3 targeting CD55. Cells were centrifuged at 500 x g for 5 min and washed with PBS. Total RNA was extracted using RNeasy Mini Kit (QIAGEN). Poly(A) mRNA was enriched using poly(A) mRNA Magnetic Isolation Module (New England Biolabs). Library preparations were carried out using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs), starting with 1000 ng total RNA. Final libraries were assessed using an Agilent 2100 TapeStation system (Agilent), and sequenced as paired end 150 base pair reads on a NovaSeq X (Illumina).