Difference between revisions of "Part:BBa K4335012"
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pCrU6.3 is a previously characterized RNA polymerase III promoter from the chromosome 8 of <i>Chlamydomonas reinhardtii</i>
 
pCrU6.3 is a previously characterized RNA polymerase III promoter from the chromosome 8 of <i>Chlamydomonas reinhardtii</i>
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<br>The first prerequisite for CRISPR/Cas9 experiments is sgRNA transcription under the control of the RNA polymerase III promoter (RNAPII) . In general, the RNAPII transcription factor binding site is intrinsic to the transcriptional sequence and therefore not amenable to sgRNA transcription. U6snRNA is one of the few examples of sgRNAs with proximal RNAPII promoter elements. Therefore, we used previously characterized U6snRNA sequences from chromosome 8.
 
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<h2>Usage</h2>
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pCrU6.3 was constructed into plasmid pTX2038 and pTX2040 as the promoter of gRNA, and was successfully introduced into Chlamydomonas reinhardtii to obtain the positive transformant.
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<h2>Discription</h2>
 
<h2>Discription</h2>
 
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Revision as of 13:29, 10 October 2022


pCrU6.3

pCrU6.3 is a previously characterized RNA polymerase III promoter from the chromosome 8 of Chlamydomonas reinhardtii

The first prerequisite for CRISPR/Cas9 experiments is sgRNA transcription under the control of the RNA polymerase III promoter (RNAPII) . In general, the RNAPII transcription factor binding site is intrinsic to the transcriptional sequence and therefore not amenable to sgRNA transcription. U6snRNA is one of the few examples of sgRNAs with proximal RNAPII promoter elements. Therefore, we used previously characterized U6snRNA sequences from chromosome 8.

Usage

pCrU6.3 was constructed into plasmid pTX2038 and pTX2040 as the promoter of gRNA, and was successfully introduced into Chlamydomonas reinhardtii to obtain the positive transformant.

Discription


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 48
    Illegal PstI site found at 121
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 48
    Illegal PstI site found at 121
    Illegal NotI site found at 453
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 48
    Illegal PstI site found at 121
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 48
    Illegal PstI site found at 121
  • 1000
    COMPATIBLE WITH RFC[1000]