RT Journal Article
SR Electronic
T1 CRISPR/Cas9 screening using unique molecular identifiers
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 114355
DO 10.1101/114355
A1 Bernhard Schmierer
A1 Sandeep K. Botla
A1 Jilin Zhang
A1 Mikko Turunen
A1 Teemu Kivioja
A1 Jussi Taipale
YR 2017
UL http://biorxiv.org/content/early/2017/05/15/114355.abstract
AB Loss of function screening by CRISPR/Cas9 gene knockout with pooled, lentiviral guide libraries is a widely applicable method for systematic identification of genes contributing to diverse cellular phenotypes. Here, random sequence labels (RSLs) are incorporated into the guide library, which act as unique molecular identifiers (UMIs) to allow massively parallel lineage tracing and lineage dropout screening. RSLs greatly improve the reproducibility of results by increasing both the precision and the accuracy of screens. They reduce the number of cells and sequencing reads needed to reach a set statistical power, or allow a more robust screen using the same number of cells.