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Related Experiment Video

Updated: Nov 14, 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

A robust method for protein depletion based on gene editing.

Yanhui Yang¹, Wenyi Wu², Tong Liu³

¹Ningxia Key Laboratory of Prevention and Control of Common Infectious Diseases, School of Basic Medical Sciences, Ningxia Medical University, Yinchuan, Ningxia, China.

Methods (San Diego, Calif.)

|March 11, 2021

Summary

This summary is machine-generated.

This study presents a simplified protocol for protein depletion using CRISPR-Cas9 gene editing technology. The method aids in understanding gene functions by enabling targeted protein knockout in cells.

Keywords:

CRISPR-Cas9 Lentivirus Mammalian cells Protein depletion

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Published on: June 16, 2017

Tuning Degradation to Achieve Specific and Efficient Protein Depletion

Tuning Degradation to Achieve Specific and Efficient Protein Depletion

Published on: July 20, 2019

Related Experiment Videos

Last Updated: Nov 14, 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

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

Tuning Degradation to Achieve Specific and Efficient Protein Depletion

Tuning Degradation to Achieve Specific and Efficient Protein Depletion

Published on: July 20, 2019

Area of Science:

Molecular Biology
Gene Editing
Cell Biology

Background:

Clustered regularly interspaced short palindromic repeats (CRISPR)-associated nuclease Cas9 (CRISPR-Cas9) is a powerful tool for genetic manipulation.
Protein depletion is crucial for understanding gene function.
Existing methods for protein depletion can be complex.

Purpose of the Study:

To present a simplified, step-by-step protocol for protein depletion using CRISPR-Cas9.
To validate the protocol's efficacy in a specific gene editing context.
To facilitate broader application of gene editing for functional genomics.

Main Methods:

Detailed protocol development for guide RNA design and vector construction.
Lentivirus production and cell line selection procedures.
Experimental validation of targeted protein depletion (PDGFRβ) in human epithelial cells.

Main Results:

Successful implementation of a standardized CRISPR-Cas9 based protein depletion protocol.
Demonstrated efficacy of the protocol through targeted editing of PDGFRβ.
The protocol enables efficient protein knockout in vitro and in vivo.

Conclusions:

The presented protocol offers a simplified and detailed approach for CRISPR-Cas9 mediated protein depletion.
This method is broadly applicable for studying specific gene functions.
The protocol will aid researchers in advancing functional genomics studies.