Part:BBa_K3286040
F. novicida dCpf1 (dCas12a) protein
Endonuclease dead version of the Francisella novicida cpf1 gene (Zetsche et al., 2015).
The Fn-dCpf1 protein can highly efficient bind target DNA, but due to mutations in the DNA cleavage domains it is 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) (Leenay et al., 2016).
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
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
Sequence and Features
- 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]
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