Difference between revisions of "Part:BBa K3286040"
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Endonuclease dead version of the Francisella novicida cpf1 gene (Zetsche et al., 2015).  
 
Endonuclease dead version of the Francisella novicida cpf1 gene (Zetsche et al., 2015).  
−
The Fn-dCpf1 protein is able to highly efficient bind target DNA, but due to mutations in 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 gene coding sequence).  
+
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).  
 
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).  
  

Revision as of 12:21, 14 October 2019


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


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 16
    Illegal PstI site found at 280
    Illegal PstI site found at 1543
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE 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
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 16
    Illegal PstI site found at 280
    Illegal PstI site found at 1543
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE WITH RFC[1000]