Kidney cancer metastasis CRISPR screening

1. Orthotopic kidney transplant experiments were attempted with UMARC2 and A498, injecting approximately 100,000 cells/50-100uL.2. Mixed verification crispr small banks were synthesized and introduced into UMARC2 and A498 cells respectively (and then stably transferred into the GFP-LUC plasmid), and the orthotopic tumor experiment was started, with 5 mice per cell. Image tracking begins when the tumor is larger than>500 mm3, focusing on the liver and lungs. 5 -6 months later, the kidneys, liver, and lungs are photographed and counted, and the whole tissue DNA is extracted, stored and sequenced together with the original tumor cell DNA.