Difference between revisions of "Part:BBa K3286040"
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Endonuclease dead version of the <em>Francisella novicida</em> cpf1 gene [1] | Endonuclease dead version of the <em>Francisella novicida</em> cpf1 gene [1] | ||
| − | The Fn-dCpf1 protein can highly efficient bind target DNA, but due to mutations in the DNA cleavage domains | + | The Fn-dCpf1 protein can highly efficient bind target DNA, but due to mutations in the DNA cleavage domains unable to cleave target DNA. |
The Fn-dCpf1 protein can be used as a transcriptional regulator for gene expression by blockage of RNA polymerase initiation (targeting a promoter region) or RNA polymerase elongation (targeting the coding sequence). | The Fn-dCpf1 protein can be used as a transcriptional regulator for gene expression by blockage of RNA polymerase initiation (targeting a promoter region) or RNA polymerase elongation (targeting the coding sequence). | ||
The DNA cleavage domains of the <em>F. novicida</em> Cpf1 protein were inactivated by the introduction of two mutations in the RuvC I and the RuvC II domains (D917A, E1006A) [2]. | The DNA cleavage domains of the <em>F. novicida</em> Cpf1 protein were inactivated by the introduction of two mutations in the RuvC I and the RuvC II domains (D917A, E1006A) [2]. | ||
Revision as of 20:02, 16 October 2019
F. novicida dCpf1 (dCas12a) protein
Endonuclease dead version of the Francisella novicida cpf1 gene [1]
The Fn-dCpf1 protein can highly efficient bind target DNA, but due to mutations in the DNA cleavage domains unable to cleave target DNA. The Fn-dCpf1 protein can be used as a transcriptional regulator for gene expression by blockage of RNA polymerase initiation (targeting a promoter region) or RNA polymerase elongation (targeting the coding sequence). The DNA cleavage domains of the F. novicida Cpf1 protein were inactivated by the introduction of two mutations in the RuvC I and the RuvC II domains (D917A, E1006A) [2].
- Zetsche, B., Gootenberg, J. S., Abudayyeh, O. O., Slaymaker, I. M., Makarova, K. S., Essletzbichler, P., … Zhang, F. (2015). Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell, 163(3), 759–771. https://doi.org/10.1016/j.cell.2015.09.038
- Leenay, R. T., Maksimchuk, K. R., Slotkowski, R. A., Agrawal, R. N., Gomaa, A. A., Briner, A. E., … Beisel, C. L. (2016). Identifying and Visualizing Functional PAM Diversity across CRISPR-Cas Systems. Molecular Cell, 62(1), 137–147. https://doi.org/10.1016/j.molcel.2016.02.031
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 16
Illegal PstI site found at 280
Illegal PstI site found at 1543 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 16
Illegal NheI site found at 1549
Illegal PstI site found at 280
Illegal PstI site found at 1543 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 16
Illegal BglII site found at 1502
Illegal BglII site found at 1536
Illegal BglII site found at 1622 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 16
Illegal PstI site found at 280
Illegal PstI site found at 1543 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 16
Illegal PstI site found at 280
Illegal PstI site found at 1543
Illegal NgoMIV site found at 3231 - 1000COMPATIBLE WITH RFC[1000]