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CRISPR

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Related Experiment Video

Updated: Feb 21, 2026

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

RNA targeting with CRISPR-Cas13.

Omar O Abudayyeh^1,2,3,4,5, Jonathan S Gootenberg^1,2,3,4,6, Patrick Essletzbichler^1,2,3,4

¹Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, USA.

|October 5, 2017

Summary

This summary is machine-generated.

Scientists engineered CRISPR-Cas13a for precise RNA manipulation in mammalian cells. This new tool offers targeted RNA knockdown and live-cell transcript tracking with improved specificity over existing methods.

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Last Updated: Feb 21, 2026

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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

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Molecular tools for RNA manipulation and measurement are limited.
Current methods like RNA interference have off-target effects.
Visualizing RNA often requires exogenous tags.

Purpose of the Study:

To engineer the CRISPR-Cas13a system for RNA knockdown and binding in mammalian cells.
To evaluate the efficacy and specificity of LwaCas13a for RNA targeting.
To develop a programmable platform for studying RNA in live cells.

Main Methods:

Screened 15 Cas13a orthologues to identify the most effective variant (LwaCas13a).
Expressed LwaCas13a heterologously in mammalian and plant cells.
Utilized catalytically inactive LwaCas13a for RNA binding and tracking.

Main Results:

LwaCas13a demonstrated effective targeted knockdown of reporter and endogenous transcripts.
Achieved knockdown levels comparable to RNA interference but with improved specificity.
Demonstrated programmable tracking of transcripts in live cells using inactive LwaCas13a.

Conclusions:

CRISPR-Cas13a is a versatile platform for RNA research in mammalian cells.
LwaCas13a offers a specific and programmable tool for RNA knockdown and visualization.
This technology has potential applications in therapeutic development for RNA-related diseases.