Part:BBa_K5036004

d-CAS9(C)

Part Description

It is a modified version of the CRISPR-Cas9 gene editing tool which cannot cut DNA. Instead, it can bind to a specific DNA sequence guided by an RNA molecule so it can be fused to transcriptional activators or repressors. In our model dcas9 was divided into N-terminal and C-terminal fragments. So the the C-terminal fragment was grafted onto NLS to generate NLS-dCas9(C) and avoid dcas9 self assembly

Usage

This part is attached to NLS and dCas9-synRTK receptor and won’t be released until cleavage of TCS1 and TCS2 This prevents the self-assembly of the two domains of dCas9 therefore providing control over the transcription activity.

literature characterization

In this study,The researchers used four strains of E. coli bacteria: DH5alpha and JM109 (both dam+/dcm+), and JM110 and GM2163 (dam-/dcm-). They introduced two plasmids, pRA-cas9 and pRA-dcas9, into each strain. These plasmids express Cas9 or dCas9 proteins under the control of a powerful, constantly active promoter called PgroESL.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 577
  Illegal NgoMIV site found at 650
  Illegal NgoMIV site found at 1135
  Illegal NgoMIV site found at 2044
  
• 1000
  COMPATIBLE WITH RFC[1000]