Difference between revisions of "Part:BBa K2148015"

 
 
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This contains an sgRNA which has 20 nucleotide complementary sequences with the Chlamydomonas reinhardtii chloroplast genome upstream of the cut-site, which is at the trnE2-psbH integenic region with psaA promoter. This can be used together with Cas9 to perform site-specific cutting. This part is coded as a Promoter/5UTR/NTAG/CDS/CTAG Phytobrick.
 
This contains an sgRNA which has 20 nucleotide complementary sequences with the Chlamydomonas reinhardtii chloroplast genome upstream of the cut-site, which is at the trnE2-psbH integenic region with psaA promoter. This can be used together with Cas9 to perform site-specific cutting. This part is coded as a Promoter/5UTR/NTAG/CDS/CTAG Phytobrick.
  
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===Usage and Biology===
 
===Usage and Biology===
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This part can be used with a system that manages to express Cas9 protein. It serves as the sgRNA which together with Cas9 can perfrom site-specific cuts on DNA that contains complementary sequences to the target RNA sequence in the sgRNA.
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<partinfo>BBa_K2148015 parameters</partinfo>
 
<partinfo>BBa_K2148015 parameters</partinfo>
 
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===Characterisation===
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===References===
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Alex Reis, Ph.D., Bitesize Bio, Breton Hornblower, Ph.D., Brett Robb, Ph.D. and George Tzertzinis, Ph.D., New England Biolabs, Inc. CRISPR/Cas9 and Targeted Genome Editing: A New Era in Molecular Biology. NEB expressions Issue I, 2014
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John G Doench, Ella Hartenian, Daniel B Graham, Zuzana Tothova, Mudra Hegde, Ian Smith, Meagan Sullender, Benjamin L Ebert, Ramnik J Xavier & David E Root. Rational design of highly active sgRNAs for CRISPR-Cas9–mediated gene inactivation. Nature Biotechnology 32, 1262–1267 (2014), doi:10.1038/nbt.3026
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Matthew H Larson, Luke A Gilbert, Xiaowo Wang, Wendell A Lim, Jonathan S Weissman & Lei S Qi. CRISPR interference (CRISPRi) for sequence-specific control of gene expression. Nature Protocols 8, 2180–2196 (2013) doi:10.1038/nprot.2013.132

Latest revision as of 19:32, 13 October 2016


gRNA for AGG PAM

This contains an sgRNA which has 20 nucleotide complementary sequences with the Chlamydomonas reinhardtii chloroplast genome upstream of the cut-site, which is at the trnE2-psbH integenic region with psaA promoter. This can be used together with Cas9 to perform site-specific cutting. This part is coded as a Promoter/5UTR/NTAG/CDS/CTAG Phytobrick.

Usage and Biology

This part can be used with a system that manages to express Cas9 protein. It serves as the sgRNA which together with Cas9 can perfrom site-specific cuts on DNA that contains complementary sequences to the target RNA sequence in the sgRNA.


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]


Characterisation

References

Alex Reis, Ph.D., Bitesize Bio, Breton Hornblower, Ph.D., Brett Robb, Ph.D. and George Tzertzinis, Ph.D., New England Biolabs, Inc. CRISPR/Cas9 and Targeted Genome Editing: A New Era in Molecular Biology. NEB expressions Issue I, 2014

John G Doench, Ella Hartenian, Daniel B Graham, Zuzana Tothova, Mudra Hegde, Ian Smith, Meagan Sullender, Benjamin L Ebert, Ramnik J Xavier & David E Root. Rational design of highly active sgRNAs for CRISPR-Cas9–mediated gene inactivation. Nature Biotechnology 32, 1262–1267 (2014), doi:10.1038/nbt.3026

Matthew H Larson, Luke A Gilbert, Xiaowo Wang, Wendell A Lim, Jonathan S Weissman & Lei S Qi. CRISPR interference (CRISPRi) for sequence-specific control of gene expression. Nature Protocols 8, 2180–2196 (2013) doi:10.1038/nprot.2013.132