Composite
crRNA-rrnB

Part:BBa_K2627004

Designed by: Zhaoqin Zhang   Group: iGEM18_SDU-CHINA   (2018-10-06)
Revision as of 12:11, 6 October 2018 by Registry (Talk | contribs)

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crRNA targeting GltA

Type I-E CRISPRi system is an endogenous CRISPRi system in Escherichia coli improved from type I-E CRISPR system. In this system, a small guide RNA(crRNA) together with Cascade protein is produced and bind to a specific DNA sequence by base complementary. Then the Cas3 protein will be recruited by Cascade and cut the DNA sequence recognized by crRNA. So the specific DNA will be removed from bacteria genome or plasmids, and this gene will no longer function. Type I-E CRISPRi system is built on the basis of Cas3 protein is knocked out from the genome, so the DNA will not be removed. However, the Cascade-crRNA complex will still prevent RNA polymerase from moving through this part, so the expression of this gene will be blocked partially, which is easier to operate. We used this part to produce crRNA that specifically recognize gltA to block the expression of gltA, thus we can block TCA cycle partially, which allows us to redirect metabolic flux to produce PHB

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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 314


[edit]
Categories
//chassis/prokaryote/ecoli
//function/crispr
Parameters
None