TY - JOUR
T1 - Improvement of the CRISPR-Cpf1 system with ribozyme-processed crRNA
AU - Gao, Zongliang
AU - Herrera-Carrillo, Elena
AU - Berkhout, Ben
PY - 2018
Y1 - 2018
N2 - 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 3ʹ-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.
AB - 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 3ʹ-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.
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85057580447&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/30470168
U2 - https://doi.org/10.1080/15476286.2018.1551703
DO - https://doi.org/10.1080/15476286.2018.1551703
M3 - Article
C2 - 30470168
SN - 1547-6286
VL - 15
SP - 1458
EP - 1497
JO - RNA biology
JF - RNA biology
IS - 12
ER -