• RNA biology · Jan 2018

    Improvement of the CRISPR-Cpf1 system with ribozyme-processed crRNA.

    • Zongliang Gao, Elena Herrera-Carrillo, and Ben Berkhout.
    • a Laboratory of Experimental Virology, Department of Medical Microbiology , Amsterdam UMC, University of Amsterdam , Amsterdam , The Netherlands.
    • RNA Biol. 2018 Jan 1; 15 (12): 1458-1467.

    AbstractThe 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.

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