Difference between revisions of "Part:BBa K1026002"
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<partinfo>BBa_K1026002 short</partinfo> | <partinfo>BBa_K1026002 short</partinfo> | ||
| − | This Constitutively Expressed gRNA targeting mRFP is a no-BioBrick-scars assembly of the constitutive promoter BBa_J23100, DNA sequence of a gRNA that targets mRFP reporter and a reliable terminator BBa_B0015. This gRNA sequence comes from the following literature: | + | This Constitutively Expressed gRNA targeting mRFP is a no-BioBrick-scars assembly of the constitutive promoter BBa_J23100, DNA sequence of a gRNA that targets mRFP reporter and a reliable terminator BBa_B0015. |
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| + | This gRNA sequence comes from the following literature: | ||
QI, LEI S., LARSON, MATTHEW H., GILBERT, LUKE A., DOUDNA, JENNIFER A., WEISSMAN, JONATHAN S., ARKIN, ADAM P. & LIM, WENDELL A. 2013. Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell, 152, 1173-1183. | QI, LEI S., LARSON, MATTHEW H., GILBERT, LUKE A., DOUDNA, JENNIFER A., WEISSMAN, JONATHAN S., ARKIN, ADAM P. & LIM, WENDELL A. 2013. Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell, 152, 1173-1183. | ||
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In CRISPRi system, when dCas9 and gRNA bind together, they will bind to the specific DNA that has the complementary sequence of gRNA. We test this system using mRFP as a reporter gene. | In CRISPRi system, when dCas9 and gRNA bind together, they will bind to the specific DNA that has the complementary sequence of gRNA. We test this system using mRFP as a reporter gene. | ||
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There are two experimenting groups: e.coli in control group only expresses dCas9 and mRFP, and in case group it expresses dCas9, sgRNA and mRFP. And the result is shown as follows: | There are two experimenting groups: e.coli in control group only expresses dCas9 and mRFP, and in case group it expresses dCas9, sgRNA and mRFP. And the result is shown as follows: | ||
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[[File:Example.jpg]] | [[File:Example.jpg]] | ||
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In case group, dCas9 did not affect the expression of mRFP; in control group, there is a significant knock down effect of mRFP. | In case group, dCas9 did not affect the expression of mRFP; in control group, there is a significant knock down effect of mRFP. | ||
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All of these results are in consistent with our expectation. | All of these results are in consistent with our expectation. | ||
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| + | <!-- Add more about the biology of this part here | ||
| + | ===Usage and Biology=== | ||
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Revision as of 17:56, 27 September 2013
Constitutively Expressed gRNA targeting mRFP
This Constitutively Expressed gRNA targeting mRFP is a no-BioBrick-scars assembly of the constitutive promoter BBa_J23100, DNA sequence of a gRNA that targets mRFP reporter and a reliable terminator BBa_B0015.
This gRNA sequence comes from the following literature:
QI, LEI S., LARSON, MATTHEW H., GILBERT, LUKE A., DOUDNA, JENNIFER A., WEISSMAN, JONATHAN S., ARKIN, ADAM P. & LIM, WENDELL A. 2013. Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell, 152, 1173-1183.
In CRISPRi system, when dCas9 and gRNA bind together, they will bind to the specific DNA that has the complementary sequence of gRNA. We test this system using mRFP as a reporter gene.
There are two experimenting groups: e.coli in control group only expresses dCas9 and mRFP, and in case group it expresses dCas9, sgRNA and mRFP. And the result is shown as follows:
In case group, dCas9 did not affect the expression of mRFP; in control group, there is a significant knock down effect of mRFP.
All of these results are in consistent with our expectation.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]