Difference between revisions of "Part:BBa K3885171"

 
 
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<partinfo>BBa_K3885171 short</partinfo>
 
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This is a special engineered gRNA that targets the middle of tetR gene, which bound to Cas9/dCas9. It also contains a RNA-responsie site in the 5' end of the igRNA, which can change their conformation to the active state exclusively upon hybridization with single-strand trigger RNAs (target RNAs).
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The rational design of igRNA is adding additional sequences in the 5' of sgRNA. <br>
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<img src="https://static.igem.org/mediawiki/parts/a/a9/3-1111.png" width=95% style="display: block;margin: 10px auto;"/>
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===Usage and Biology===
 
===Usage and Biology===
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<p style="text-align: center;"> Figure 1. Schematic of gRNA and igRNA.</p>
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The “igRNA” stands for RNA-interacting guide RNA, which switch to their active state only upon interaction with target RNAs (Figure 1).<br>
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The additional sequences include recognition sequence A, cis-repressed sequence and recognition sequence B. The cis-repressed sequence is complementary to the spacer, leading to the repression of the binding ability between igRNA with its DNA target. The recognition sequence A and the recognition sequence B are complementary to the interacting sequence A and interacting sequence B, respectively.<br>
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All designed igRNAs were analyzed using <a href="http://www.nupack.org/">NUPACK webserver</a>.<br>
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The igRNA that has lower free energy, co-folding energy with target RNA as well as reduced self-comlementarity at 29℃ would be the best (Figure 2).
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Latest revision as of 14:32, 20 October 2021


Interacting Guide RNA（igRNA）

The rational design of igRNA is adding additional sequences in the 5' of sgRNA.

Usage and Biology

Figure 1. Schematic of gRNA and igRNA.

The “igRNA” stands for RNA-interacting guide RNA, which switch to their active state only upon interaction with target RNAs (Figure 1).
The additional sequences include recognition sequence A, cis-repressed sequence and recognition sequence B. The cis-repressed sequence is complementary to the spacer, leading to the repression of the binding ability between igRNA with its DNA target. The recognition sequence A and the recognition sequence B are complementary to the interacting sequence A and interacting sequence B, respectively.

All designed igRNAs were analyzed using NUPACK webserver.
The igRNA that has lower free energy, co-folding energy with target RNA as well as reduced self-comlementarity at 29℃ would be the best (Figure 2).


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
    COMPATIBLE WITH RFC[1000]