Difference between revisions of "Part:BBa K2627004"
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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. | 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. | ||
| − | [[File:T--SDU-China--crrna.png| | + | [[File:T--SDU-China--crrna.png|500px|center]] |
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===Usage and Biology=== | ===Usage and Biology=== | ||
Revision as of 15:58, 17 October 2018
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 314