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Updated: Nov 11, 2025

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

CRISPR/Cas9-based directed evolution in mammalian cells.

Oliver Griesbeck¹

¹Max-Planck-Institut für Neurobiologie, Am Klopferspitz 18, 82152, Martinsried, Germany.

Current Opinion in Structural Biology

|March 30, 2021

Summary

This summary is machine-generated.

New CRISPR/Cas-based methods enable protein directed evolution in mammalian cells. This advances protein engineering for systems where the cellular environment is crucial.

Keywords:

Base editor CRISPR Cassette mutagenesis Protein engineering Screening

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Published on: February 26, 2019

Area of Science:

Biochemistry
Molecular Biology
Protein Engineering

Background:

Traditional directed evolution methods often utilize in vitro techniques or microbial systems.
Mammalian cellular environments are critical for studying and engineering certain protein classes.

Purpose of the Study:

To introduce novel CRISPR/Cas-based strategies for protein diversification within mammalian cells.
To explore in situ protein evolution approaches in a native cellular context.

Main Methods:

CRISPR/Cas9-mediated indel mutagenesis for protein diversification.
Homology-directed repair for cassette mutagenesis of protein-coding sequences.
Fusions of Cas9 variants with base editors and other effectors to introduce targeted mutations.

Main Results:

Demonstration of effective protein diversification using CRISPR/Cas-based tools in mammalian cells.
Establishment of in situ evolution methods as a viable alternative to traditional approaches.
Expansion of the toolkit for engineering proteins requiring a mammalian cellular environment.

Conclusions:

CRISPR/Cas-based directed evolution in mammalian cells represents a significant advancement in protein engineering.
These methods offer powerful new avenues for discovering and optimizing proteins in relevant biological contexts.
The mammalian cellular context is increasingly accessible for sophisticated protein evolution studies.