Difference between revisions of "Part:BBa K1062001"
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<partinfo>BBa_K1062001 short</partinfo> | <partinfo>BBa_K1062001 short</partinfo> | ||
| − | This part is a guide RNA (gRNA) that targets GFP, when bound to Cas9. It also contains a cleavage site recognized by the Csy4 enzyme which allows it to be cleaved from neighboring DNA/RNA. | + | This part is a guide RNA (gRNA) that targets the GFP gene, when bound to Cas9. It also contains a cleavage site recognized by the Csy4 enzyme which allows it to be cleaved from neighboring DNA/RNA. |
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In both of our projects, we used gRNAs and dCas9 as: | In both of our projects, we used gRNAs and dCas9 as: | ||
| − | 1) To specifically target and repress the "virulence gene" (we used flourescent proteins as a proof of concept) in our conjugation project. However, the gRNA for GFP is not used in the conjugation project since the green fluorescent protein | + | 1) To specifically target and repress the "virulence gene" (we used flourescent proteins as a proof of concept) in our conjugation project. However, the gRNA for GFP is not used in the conjugation project since the green fluorescent protein was used to represent our donor strain of the project. |
2) Improve past circuits with better decision making by using dCas9 instead of traditional repressors. | 2) Improve past circuits with better decision making by using dCas9 instead of traditional repressors. | ||
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[[File:Synthetic Circuit.jpg]] | [[File:Synthetic Circuit.jpg]] | ||
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| + | In the Synthetic Circuit, the target is the GFP gene that would be located in the '''Low Concentration Pathway'''. The purpose of the gRNA is to prevent any possibility of the GFP gene expressing, due to a possible leaky promoter, when the '''High Concentration Pathway''' is being induced. | ||
<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
Latest revision as of 09:08, 24 September 2013
Guide RNA (gRNA) target for GFP
This part is a guide RNA (gRNA) that targets the GFP gene, when bound to Cas9. It also contains a cleavage site recognized by the Csy4 enzyme which allows it to be cleaved from neighboring DNA/RNA.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
In both of our projects, we used gRNAs and dCas9 as:
1) To specifically target and repress the "virulence gene" (we used flourescent proteins as a proof of concept) in our conjugation project. However, the gRNA for GFP is not used in the conjugation project since the green fluorescent protein was used to represent our donor strain of the project.
2) Improve past circuits with better decision making by using dCas9 instead of traditional repressors.
Conjugation Project
The GFP gRNA was not used in the Conjugation Project
Synthetic Circuit
In the Synthetic Circuit, the target is the GFP gene that would be located in the Low Concentration Pathway. The purpose of the gRNA is to prevent any possibility of the GFP gene expressing, due to a possible leaky promoter, when the High Concentration Pathway is being induced.