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An integrated pipeline for mammalian genetic screening.

ABSTRACT: 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.

SUBMITTER: Kramme C

PROVIDER: S-EPMC9017118 | biostudies-literature | 2021 Oct

REPOSITORIES: biostudies-literature

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An integrated pipeline for mammalian genetic screening.

Kramme Christian C Plesa Alexandru M AM Wang Helen H HH Wolf Bennett B Smela Merrick Pierson MP Guo Xiaoge X Kohman Richie E RE Chatterjee Pranam P Church George M GM

Cell reports methods 20210927 6

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 whi ...[more]

PMID: 35474898

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