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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

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Published on: June 16, 2017

Genetic Engineering and Screening Using Base Editing and Inducible Gene Knockout.

Esther Feng Ying Ng¹, Franz Meitinger²

¹Okinawa Institute of Science and Technology Graduate University, Okinawa, Japan.

Methods in Molecular Biology (Clifton, N.J.)

|December 1, 2024

Summary

This study presents methods for creating gene editing tools, specifically base editors and inducible knockout cell lines. These tools enable precise genetic screening to understand gene and protein functions in human cells.

Keywords:

Base editing CRISPR Cas9 Gene editing Gene engineering Gene knockout Genetic screening

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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

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Published on: April 11, 2019

Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

Genetic screens are crucial for identifying gene functions and drug responses.
Base editors allow targeted single amino acid mutations, enhancing functional domain analysis.

Purpose of the Study:

To outline methods for generating efficient base editor and inducible knockout cell lines.
To enable targeted gene manipulation for genetic screens.

Main Methods:

Development of base editor cell lines for targeted mutagenesis.
Establishment of inducible knockout cell lines for gene function studies.
Application of these tools in genetic screens for gene and domain function analysis.

Main Results:

Efficient creation of base editor and inducible knockout cell lines.
Demonstration of their utility in targeted genetic screens.
Elucidation of gene and protein domain functions within cellular contexts.

Conclusions:

These methods provide powerful tools for precise gene function analysis.
Facilitates the discovery of essential genes and drug response mechanisms.
Advances the understanding of gene and domain roles in cell biology.