GSM9090499: LLC, Rag1-/-, rep12; Mus musculus; OTHER
Tumors were harvested when they reached approximately 1000 mm3 in size. The QIAamp DNA Mini Kit (Qiagen) was used for genomic DNA extraction, followed by PCR for sgRNA amplification and NGS sequencing. Using RNA-seq data from 22 patients, we identified DEGs associated with immunotherapy efficacy through edgeR. A total of 394 genes were significantly downregulated in DCR patients, with |LogFC|>1, p-value<0.05. These human genes were subsequently converted to their mouse orthologs, resulting in 332 mouse genes. For each gene, 6-10 sgRNAs were designed, culminating in a customized CRISPR library comprising 3689 sgRNAs.