Cpf1

from Wikipedia, the free encyclopedia
CRISPR-associated endonuclease Cpf1
other names
  • FnCpf1
  • Cas12a

Existing structure data : PDB  5MGA , PDB  5NFV , PDB  5NG6

Mass / length primary structure 1,300 amino acids , 151,915 Da
Identifier
External IDs
Enzyme classification
EC, category 3.1.-.-
Orthologue
Francisella novicida Prevotella ruminicola
Entrez NV NV
UniProt A0Q7Q2 A0A1M7G246
PubMed search NV NV

Cpf1 (from CRISPR-associated endonuclease in Prevotella and Francisella 1 , also Cas12a ) is an endonuclease and a ribonucleoprotein from the antiviral defense CRISPR in the bacterial genera Prevotella and Francisella .

properties

Cpf1 is an endonuclease that a single-g RNA binds, and then a complementary DNA to the gRNA binds and cuts. In addition, Cpf1 also cuts gRNA precursors to form active gRNA. If Cpf1 in E. coli expressed , will be plasmids with homologous DNA sequences cut for the first CRISPR spacer and degraded. Cpf1 cuts dsDNA and creates a sticky end with a 5 'overhang of four or five nucleotides at a distance between 18 or 22 to 23 nucleotides after the Protospacer Adjacent Motif with the sequence 5'- T TTV-3' (where V stands for C , G or A ). In addition, C-containing PAMs with a different sequence are also bound, but with less efficiency.

Applications

The Cpf1 is used in the CRISPR / Cpf1 system for genome editing . By cutting gRNA, several different gRNAs can be generated from one mRNA , which facilitates the simultaneous cutting of several DNA sequences ( multiplex genome editing ). Various variants of Cpf1 that bind other PAMs have been generated through protein engineering .

Web links

Individual evidence

  1. ↑ a b c B. Zetsche, JS Gootenberg, OO Abudayyeh, IM Slaymaker, KS Makarova, P. Essletzbichler, SE Volz, J. Joung, J. van der Oost, A. Regev, EV Koonin, F. Zhang: Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. In: Cell. Volume 163, number 3, October 2015, pp. 759-771, doi : 10.1016 / j.cell.2015.09.038 , PMID 26422227 , PMC 4638220 (free full text).
  2. ↑ DC Swarts, M. Jinek: Cas9 versus Cas12a / Cpf1: Structure-function comparisons and implications for genome editing. In: Wiley Interdiscip Rev RNA. May 2018, 22: e1481., Doi : 10.1002 / wrna.1481 , PMID 29790280 .
  3. ↑ T. Yamano, B. Zetsche, R. Ishitani, F. Zhang, H. Nishimasu, O. Nureki: Structural Basis for the Canonical and Non-canonical PAM Recognition by CRISPR-Cpf1. In: Molecular cell. Volume 67, number 4, August 2017, pp. 633–645.e3, doi : 10.1016 / j.molcel.2017.06.035 , PMID 28781234 .
  4. ↑ G. Zhong, H. Wang, Y. Li, MH Tran, M. Farzan: Cpf1 proteins excise CRISPR RNAs from mRNA transcripts in mammalian cells. In: Nature chemical biology. Volume 13, number 8, August 2017, pp. 839-841, doi : 10.1038 / nchembio.2410 , PMID 28628097 , PMC 5577360 (free full text).
  5. ↑ H. Nishimasu, T. Yamano, L. Gao, F. Zhang, R. Ishitani, O. Nureki: Structural Basis for the Altered PAM Recognition by Engineered CRISPR-Cpf1. In: Molecular cell. Volume 67, number 1, July 2017, pp. 139-147.e2, doi : 10.1016 / j.molcel.2017.04.019 , PMID 28595896 .
  6. ^ L. Gao, DB Cox, WX Yan, JC Manteiga, MW Schneider, T. Yamano, H. Nishimasu, O. Nureki, N. Crosetto, F. Zhang: Engineered Cpf1 variants with altered PAM specificities. In: Nature Biotechnology . Volume 35, number 8, August 2017, pp. 789-792, doi : 10.1038 / nbt.3900 , PMID 28581492 , PMC 5548640 (free full text).