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Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...

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Related Experiment Video

Updated: Nov 26, 2025

One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster

One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster

Published on: January 26, 2024

A Simple and Efficient CRISPR Technique for Protein Tagging.

Fanning Zeng¹, Valerie Beck¹, Sven Schuierer¹

¹Novartis Institutes for Biomedical Research, 4002 Basel, Switzerland.

|December 9, 2020

Summary

This summary is machine-generated.

Homology independent gene Tagging (HiTag) offers an efficient CRISPR-based method for protein tagging in cells. This technique significantly improves genetic knock-in efficiency compared to traditional approaches, enabling broader applications in cell biology research.

Keywords:

CRISPR knock in non-homologous end-joining (NHEJ)protein tagging

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Last Updated: Nov 26, 2025

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Published on: December 24, 2016

Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

Homology-directed repair (HDR) for genetic knock-in is often inefficient.
Current methods require selection of modified cells, limiting primary cell applications.

Purpose of the Study:

To develop a novel, highly efficient method for CRISPR-mediated protein tagging.
To overcome the limitations of HDR in genetic modification of cells.

Main Methods:

Introduction of Homology independent gene Tagging (HiTag) system.
CRISPR-Cas9 mediated gene tagging in various cell types.
Electroporation for efficient delivery of reagents.

Main Results:

HiTag achieved protein tagging in up to 66% of transfected cells.
Demonstrated effectiveness across diverse cell types, including primary cells.
Successfully applied HiTag for fluorescent protein knock-in, protein localization studies, and luciferase assays.

Conclusions:

HiTag provides a robust and efficient alternative to HDR for genetic knock-in.
The method facilitates live cell imaging, protein localization, and protein level monitoring.
HiTag expands the utility of CRISPR technology in primary cells and various research applications.