GSM9192238: scPerturb_Aged3; Mus musculus; RNA-Seq
Fifteen days after tumor inoculation, ametrine⁺ CD45.1 Cas9⁺ OT-1 CD8⁺ T cells were isolated from tumors and tumor-draining lymph nodes (tdLNs) of young and aged mice using a BD FACSAria™ Fusion sorter. Sorted cells from independent mice were pooled into two biological replicates per group, counted, and loaded onto a Chromium Controller (10x Genomics) for a target recovery of 10,000 single cells per run. Libraries were prepared with the 10x Single Cell 5′ Reagent Kits v2 (Dual Index) and Feature Barcode technology for CRISPR screening, following the manufacturer's protocol. Sequencing was performed on an Illumina NovaSeq X or NextSeq 1000/2000 system using 100-cycle kits with paired-end reads. Library was performed according to the manufacter's instructions (10x Single Cell 5′ Reagent Kits v2, CG00511 RevD, 10x Genomics).