2019 •
Protocols for CRISPR-Cas9 Screening in Lymphoma Cell Lines
Authors:
Dan E. Webster, Sandrine Roulland, Sandrine Roulland, James D. Phelan
Abstract:
Genome-wide screens are a powerful technique to dissect the complex network of genes regulating diverse cellular phenotypes. The recent adaptation of the CRISPR-Cas9 system for genome engineering has revolutionized functional genomic screening. Here, we present protocols used to introduce Cas9 into human lymphoma cell lines, produce high-titer lentivirus of a genome-wide sgRNA library, transduce and culture cells during the screen, isolate genomic DNA, and prepare a custom library for next-generation sequencing. These protocols were tailored fo (...)
Genome-wide screens are a powerful technique to dissect the complex network of genes regulating diverse cellular phenotypes. The recent adaptation of the CRISPR-Cas9 system for genome engineering has revolutionized functional genomic screening. Here, we present protocols used to introduce Cas9 into human lymphoma cell lines, produce high-titer lentivirus of a genome-wide sgRNA library, transduce and culture cells during the screen, isolate genomic DNA, and prepare a custom library for next-generation sequencing. These protocols were tailored for loss-of-function CRISPR screens in human lymphoma cell lines but are highly amenable for other experimental purposes. (Read More)
Daniel E. Webster, Sandrine Roulland, James D. Phelan
Methods in Molecular Biology ·
2019
Computational biology |
Genetics |
We have placed cookies on your device to help make this website and the services we offer better. By using this site, you agree to the use of cookies. Learn more