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Related Experiment Video

Updated: Apr 21, 2026

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

Published on: December 11, 2020

Precise, specific gene editing via a compact GoCas12m-FokI chimeric nuclease.

Tin Marsic¹, Sivakrishna Rao Gundra¹, Mustapha Aouida²

¹Laboratory for Genome Engineering and Synthetic Biology, Division of Biomedical Sciences, 4700 King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.

Nucleic Acids Research

|April 20, 2026

Summary

This summary is machine-generated.

A new gene editing tool, GoCas12m-FokI, offers precise genome editing with high efficiency and no detectable off-target activity. Its compact size facilitates delivery for in vivo gene therapy applications.

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Last Updated: Apr 21, 2026

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

Published on: December 11, 2020

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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Published on: May 25, 2018

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

Area of Science:

Biotechnology
Genomics
Molecular Biology

Background:

CRISPR gene editing advanced functional genomics but faces challenges in delivery and specificity.
Therapeutic translation of gene editing is hindered by limited delivery capacity and off-target mutations.

Purpose of the Study:

To develop a novel, compact, and highly specific gene editing platform for therapeutic applications.
To engineer a chimeric system combining a guiding module with a nuclease for precise genome modification.

Main Methods:

Fused a catalytically inactive Cas12m guiding module (GoCas12m) with the FokI nuclease domain to create the GoCas12m-FokI editor.
Tested the editor's activity on surrogate reporters and endogenous human loci (CLTA1, HBB, AIFM1, ABL).
Confirmed specificity using targeted deep sequencing to detect off-target activity.

Main Results:

The GoCas12m-FokI editor demonstrated robust and high-efficiency editing at clinically relevant targets.
No detectable off-target activity was observed at in silico-predicted sites.
The editor's compact size (nearly half of Cas9/Cas12a) facilitates AAV-mediated delivery.

Conclusions:

The GoCas12m-FokI system provides a precise and specific genome editing tool with a compact architecture.
This editor is a promising alternative for therapeutic genome editing and in vivo gene therapy.
The modularity and specificity of GoCas12m-FokI enhance its potential for clinical translation.