Improvement of the CRISPR-Cpf1 System with Ribozyme-processed CrRNA

Overview

Journal RNA Biol

Specialty Molecular Biology

Date 2018 Nov 25

PMID 30470168

Citations 24

Authors

Zongliang Gao

Elena Herrera-Carrillo

Ben Berkhout

Affiliations

Soon will be listed here.

Abstract

The recently discovered clustered regularly interspaced short palindromic repeats (CRISPR)-Cpf1 system expands the genome editing toolbox. This system exhibits several distinct features compared to the widely used CRISPR-Cas9 system, but has reduced gene editing efficiency. To optimize the CRISPR-Cpf1 (Cas12a) system, we report the inclusion of self-cleaving ribozymes that facilitate processing of the crRNA transcript to produce the precise guide molecule. Insertion of the 3'-terminal HDV ribozyme boosted the gene editing activity of the CRISPR-Cpf1 system ranging from 1.1 to 5.2 fold. We also demonstrate that this design can enhance CRISPR-based gene activation. We thus generated an improved CRISPR-Cpf1 system for more efficient gene editing and gene regulation.

Citing Articles

Efficient Genome Editing Using 'NanoMEDIC' AsCas12a-VLPs Produced with Pol II-Transcribed crRNA.

Borovikova S, Shepelev M, Mazurov D, Kruglova N Int J Mol Sci. 2024; 25(23).

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Strategies for improving the genome-editing efficiency of class 2 CRISPR/Cas system.

Wang L, Han H Heliyon. 2024; 10(19):e38588.

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Structure of Fanzor2 reveals insights into the evolution of the TnpB superfamily.

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CRISPR/Cas12a toolbox for genome editing in .

Zhu P, Somvanshi T, Bao J, Scheller S Front Microbiol. 2023; 14:1235616.

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Optimizing ErCas12a for efficient gene editing in Arabidopsis thaliana.

Pietralla J, Capdeville N, Schindele P, Puchta H Plant Biotechnol J. 2023; 22(2):401-412.

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