Difference between revisions of "Part:BBa K5036004"

 
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<partinfo>BBa_K5036004 short</partinfo>
 
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==Part Description==
 
It is a modified version of the CRISPR-Cas9 gene editing tool which cannot cut DNA. Instead, it can bind to a specific DNA sequence guided by an RNA molecule so it can be fused to transcriptional activators or repressors. In our model dcas9 was divided into N-terminal and C-terminal fragments. So the the C-terminal fragment was grafted onto NLS to generate NLS-dCas9(C) and avoid dcas9 self assembly
 
It is a modified version of the CRISPR-Cas9 gene editing tool which cannot cut DNA. Instead, it can bind to a specific DNA sequence guided by an RNA molecule so it can be fused to transcriptional activators or repressors. In our model dcas9 was divided into N-terminal and C-terminal fragments. So the the C-terminal fragment was grafted onto NLS to generate NLS-dCas9(C) and avoid dcas9 self assembly
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==Usage==
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This part is attached to NLS and  DocTAR receptor and won’t be released until cleavage of NCS1 and NCS2
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This prevents the self-assembly of the two domains of dCas9 therefore providing control over the transcription activity.
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==literature characterization==
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In this study,The researchers used four strains of E. coli bacteria: DH5alpha and JM109 (both dam+/dcm+), and JM110 and GM2163 (dam-/dcm-). They introduced two plasmids, pRA-cas9 and pRA-dcas9, into each strain. These plasmids express Cas9 or dCas9 proteins under the control of a powerful, constantly active promoter called PgroESL.
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Revision as of 17:37, 9 September 2024


d-CAS9(C)

Part Description

It is a modified version of the CRISPR-Cas9 gene editing tool which cannot cut DNA. Instead, it can bind to a specific DNA sequence guided by an RNA molecule so it can be fused to transcriptional activators or repressors. In our model dcas9 was divided into N-terminal and C-terminal fragments. So the the C-terminal fragment was grafted onto NLS to generate NLS-dCas9(C) and avoid dcas9 self assembly

Usage

This part is attached to NLS and DocTAR receptor and won’t be released until cleavage of NCS1 and NCS2 This prevents the self-assembly of the two domains of dCas9 therefore providing control over the transcription activity.

literature characterization

In this study,The researchers used four strains of E. coli bacteria: DH5alpha and JM109 (both dam+/dcm+), and JM110 and GM2163 (dam-/dcm-). They introduced two plasmids, pRA-cas9 and pRA-dcas9, into each strain. These plasmids express Cas9 or dCas9 proteins under the control of a powerful, constantly active promoter called PgroESL.


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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 577
    Illegal NgoMIV site found at 650
    Illegal NgoMIV site found at 1135
    Illegal NgoMIV site found at 2044
  • 1000
    COMPATIBLE WITH RFC[1000]