An integrated pipeline for mammalian genetic screening.

Journal Article (Journal Article)

With the recent advancements in genome editing, next-generation sequencing (NGS), and scalable cloning techniques, scientists can now conduct genetic screens at unprecedented levels of scale and precision. With such a multitude of technologies, there is a need for a simple yet comprehensive pipeline to enable systematic mammalian genetic screening. In this study, we develop unique algorithms for target identification and a toxin-less Gateway cloning tool, termed MegaGate, for library cloning which, when combined with existing genetic perturbation methods and NGS-coupled readouts, enable versatile engineering of relevant mammalian cell lines. Our integrated pipeline for sequencing-based target ascertainment and modular perturbation screening (STAMPScreen) can thus be utilized for a host of cell state engineering applications.

Full Text

Duke Authors

Cited Authors

  • Kramme, C; Plesa, AM; Wang, HH; Wolf, B; Smela, MP; Guo, X; Kohman, RE; Chatterjee, P; Church, GM

Published Date

  • October 2021

Published In

Volume / Issue

  • 1 / 6

Start / End Page

  • 100082 -

PubMed ID

  • 35474898

Pubmed Central ID

  • PMC9017118

Electronic International Standard Serial Number (EISSN)

  • 2667-2375

International Standard Serial Number (ISSN)

  • 2667-2375

Digital Object Identifier (DOI)

  • 10.1016/j.crmeth.2021.100082

Language

  • eng