CRISPR / Cpf1 method
The CRISPR / Cpf1 method ( English Clustered Regularly Inter Spaced Short Palindromic repeats from Prevotella and Francis Ella ) is a biochemical method for the production of genetically modified organisms .
properties
The CRISPR / Cpf1 system comes from - like the CRISPR / Cas9 system and the CRISPR / Cas12b system - an adaptive antiviral defense mechanism, the CRISPR , but from the genera Prevotella and Francisella . As with this one, methods for using the system as a tool for biotechnological research for various organisms have been and are being developed on the basis of the system.
Comparison with Cas9
Compared to the CRISPR / Cas system, the endonuclease Cpf1 (also known as Cas12a) used in the CRISPR / Cpf1 system is a smaller enzyme that only requires an RNA as a template (no tracrRNA ) to cut double-stranded DNA at a specific point. Furthermore, Cpf1 has a different interface and creates a base overhang on one of the two DNA strands ( sticky end ), which facilitates ligation . Due to the sticky-end overhang created, the orientation of a DNA to be inserted can be controlled with Cpf1, while with Cas9 the inserted DNA has a wrong orientation in half of the insertions. As for various applications of the CRISPR / Cas system, patents have also been applied for for the Cpf1-based genome manipulations .
Differences between Cas9, Cpf1 and Cas12b
| property | Cas9 | Cpf1 (Cas12a) | Cas12b |
|---|---|---|---|
| structure | 2 RNA required | 1 RNA required (no tracrRNA ) | 2 RNA required |
| Overhang | none (blunt end) | sticky end | sticky end |
| interface | proximal to the recognition sequence | distal to the recognition sequence | distal to the recognition sequence |
| Target sequence | G-rich PAM | T-rich PAM | T-rich PAM |
| Cell type | dividing cells | resting cells | unknown |
Web links
- Heidi Ledford: Tool of genetic manipulation - genetic engineering: CRISPR changes everything . In: Spectrum of Science . from June 24, 2015.
Individual evidence
- ^ R. Sorek, V. Kunin, P. Hugenholtz: CRISPR - a widespread system that provides acquired resistance against phages in bacteria and archaea. In: Nature reviews. Microbiology. Volume 6, Number 3, March 2008, pp. 181-186, ISSN 1740-1534 , doi : 10.1038 / nrmicro1793 , PMID 18157154 .
- ↑ a b Winston X. Yan, Pratyusha Hunnewell, Lauren E. Alfonse, Jason M. Carte, Elise Keston-Smith, Shanmugapriya Sothiselvam, Anthony J. Garrity, Shaorong Chong, Kira S. Makarova, Eugene V. Koonin, David R. Cheng, David A. Scott: Functionally diverse type V CRISPR-Cas systems. In: Science. 363, 2019, p. 88, doi : 10.1126 / science.aav7271 .
- ↑ a b c d Heidi Ledford: Alternative CRISPR system could improve genome editing. In: Nature. 526, 2015, p. 17, doi : 10.1038 / nature.2015.18432 .
- ^ Even CRISPR: A new way to edit DNA may speed the advance of genetic engineering . In: Economist , October 3, 2015.
- ↑ EPO grants CRISPR patent to Broad. (No longer available online.) In: transkript.de. February 28, 2017, archived from the original on March 12, 2017 ; accessed on March 9, 2017 . Info: The archive link was inserted automatically and has not yet been checked. Please check the original and archive link according to the instructions and then remove this notice.
- ↑ cas12b - CRISPR-associated endonuclease Cas12b - Alicyclobacillus acidoterrestris (strain ATCC 49025 / DSM 3922 / CIP 106132 / NCIMB 13137 / GD3B) - cas12b gene. In: uniprot.org. October 16, 2013, accessed January 24, 2019 .
