A simplified transposon mutagenesis method to perform phenotypic forward genetic screens in cultured cells.

ABSTRACT:

Background

The introduction of genome-wide shRNA and CRISPR libraries has facilitated cell-based screens to identify loss-of-function mutations associated with a phenotype of interest. Approaches to perform analogous gain-of-function screens are less common, although some reports have utilized arrayed viral expression libraries or the CRISPR activation system. However, a variety of technical and logistical challenges make these approaches difficult for many labs to execute. In addition, genome-wide shRNA or CRISPR libraries typically contain of hundreds of thousands of individual engineered elements, and the associated complexity creates issues with replication and reproducibility for these methods.

Results

Here we describe a simple, reproducible approach using the SB transp

SUBMITTER: Feddersen CR 

PROVIDER: S-EPMC6580595 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature