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Updated: Aug 25, 2025

Electroporation-Based CRISPR-Cas9-Mediated Gene Knockout in THP-1 Cells and Single-Cell Clone Isolation

Electroporation-Based CRISPR-Cas9-Mediated Gene Knockout in THP-1 Cells and Single-Cell Clone Isolation

Published on: February 28, 2025

Selecting for CRISPR-Edited Knock-In Cells.

Nina Reuven¹, Yosef Shaul¹

¹Department of Molecular Genetics, Weizmann Institute of Science, Rehovot 76100, Israel.

International Journal of Molecular Sciences

|October 14, 2022

Summary

This summary is machine-generated.

CRISPR genome editing is powerful but knock-in mutagenesis is inefficient. This review discusses strategies to select successfully edited cells, improving efficiency for research and medicine.

Keywords:

CRISPR HDR-dependent editing co-editing genome editing scarless selection

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Published on: April 11, 2019

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Last Updated: Aug 25, 2025

Electroporation-Based CRISPR-Cas9-Mediated Gene Knockout in THP-1 Cells and Single-Cell Clone Isolation

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Published on: February 28, 2025

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

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

Published on: June 16, 2017

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

CRISPR-Cas9 enables efficient genome targeting but precise mutation introduction is challenging.
Endogenous DNA repair mechanisms limit the efficiency of knock-in mutagenesis.
Current methods often result in a low percentage of successfully edited cells.

Purpose of the Study:

To review strategies for enhancing specific genome editing in mammalian cells.
To focus on methods for selecting successfully edited cells.
To improve the efficiency of targeted genetic modifications.

Main Methods:

Discussing strategies that link selectable genes to genes of interest (GOI).
Explaining co-editing techniques where GOI is edited alongside a selectable gene.
Highlighting "scarless" selection methods that modify only the GOI.

Main Results:

Various selection strategies significantly enrich populations of edited cells.
Linked selectable genes and co-editing improve the yield of desired mutations.
"Scarless" methods minimize unintended genomic alterations.

Conclusions:

Efficient selection of edited cells is crucial for advancing CRISPR applications in research and medicine.
Optimized selection strategies increase the utility of genome editing tools.
Future directions may involve further refining scarless and co-editing techniques.