Part:BBa_K5036029

dCas9(C)_NLS-Syn-VEGFR-1 (VEGF-R1, C-TEV, NLS, TCS(Q,L), HA,dCas9(C),VP64,GFP)

Part Description

In our first receptor chain, we've engineered a system that responds to tissue injury. An external domain, VEGF-R1, is attached to an internal domain composed of C terminal domain of TEV protease, a nuclear localization signal (NLS), a TEV cleavage site(TCS(Q,L)), and dCas9(C) which is linked to transcription activator VP64

Usage

this is our receptor's first chain. our receptor is activated after binding of VEGF to the external domain which is designed to attach specifically to it. after activation the two domains of TEV dimerizes forming catalytically active TEV protease which will cleave the two chains at TCS. upon cleavage of the two chains the two domains of dCas9 dimerize and is released attached to transcription activator to be guided to its direction

This figure illustrates variant of our receptor's first chain where TCS (Q, L) is to it.

Software Characterization

we had the chance to match the external domains with different internal domain components to select suitable chains. The whole chain affinity is affected by the internal domain, thus we had to try:

VEGFR1Cdcas-VEGFA

The interaction between the chain composed of VEGFR1 as an external domain and Cdcas9 as an internal domain with VEGFA yields ΔG of -11.2 kcal mol-1 .

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Unknown
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Unknown
• 25
  INCOMPATIBLE WITH RFC[25]

  Unknown
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 6737
  Illegal SapI.rc site found at 3268