Difference between revisions of "Part:BBa K3885172"

 
 
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This is a gRNA that specifically targets the middle of tetR gene&#65292;when bound to Cas9.
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<img src="https://static.igem.org/mediawiki/parts/d/da/1-11.png" width=95% style="display: block;margin: 10px auto;"/>
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Guide RNA (gRNA) is a piece of RNAs that function as guides for RNA- or DNA-targeting enzymes, which they form complexes with. Very often these enzymes will delete, insert or otherwise alter the targeted DNA. <br>
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===Usage and Biology===
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<img src="https://static.igem.org/mediawiki/parts/c/c7/1-121.png" width=95% style="display: block;margin: 10px auto;"/>
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<p style="text-align: center;"> Figure 1. Schematic of the design procedure of the gRNA construction and screening. There are five 20nt sequences to change into five different gRNAs.</p>
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In the CRISPR/Cas9 system, gRNA can direct Cas9 protein to cut dsDNA at the sequence target site. Simultaneously, a hybridization region of igRNA is adding additional sequences in the 5' of sgRNA. Therefore, designing an active gRNA with high cleavage ability is important.<br>
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Based on the target gene(tetR), we chose CGG and AGG as PAM, and designed 5 gRNAs in the plasmid pTargetF (Figure 1). <br>
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Latest revision as of 12:20, 20 October 2021


Guide RNA (gRNA) target for tetR


Guide RNA (gRNA) is a piece of RNAs that function as guides for RNA- or DNA-targeting enzymes, which they form complexes with. Very often these enzymes will delete, insert or otherwise alter the targeted DNA.

Usage and Biology

Figure 1. Schematic of the design procedure of the gRNA construction and screening. There are five 20nt sequences to change into five different gRNAs.

In the CRISPR/Cas9 system, gRNA can direct Cas9 protein to cut dsDNA at the sequence target site. Simultaneously, a hybridization region of igRNA is adding additional sequences in the 5' of sgRNA. Therefore, designing an active gRNA with high cleavage ability is important.
Based on the target gene(tetR), we chose CGG and AGG as PAM, and designed 5 gRNAs in the plasmid pTargetF (Figure 1).


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]