Book Chapter
A comprehensive protocol resource for performing pooled shRNA and CRISPR screens
LA Cluse, I Nikolic, D Knight, PB Madhamshettiwar, J Luu, KJ Cowley, T Semple, GM Arnau, J Shortt, RW Johnstone, KJ Simpson
Methods in Molecular Biology | Methods in Molecular Biology | HUMANA PRESS INC | Published : 2018
Abstract
This chapter details a compendium of protocols that collectively enable the reader to perform a pooled shRNA and/or CRISPR screen—with methods to identify and validate positive controls and subsequent hits; establish a viral titer in the cell line of choice; create and screen libraries, sequence strategies, and bioinformatics resources to analyze outcomes. Collectively, this provides an overarching resource from the start to finish of a screening project, making this technology possible in all laboratories.