Difference between revisions of "Part:BBa K3044009"

Latest revision as of 01:23, 22 October 2019

dCas9 with IPTG inducible promotor

This dCas9 is a codon optimized version of BBa_K1150000 for expression in E. coli. Together with a single guide RNA (sgRNA), the protein can be used for silencing of specific target genes. dCas9 is the inactive form of the endonuclease Cas9. In contrast to Cas9, dCas9 does not cleave the DNA. Instead, it sterically blocks the elongation of transcription as a result of the sgRNA-DNA interaction. This will only repress the expression of the target gene thus avoiding the destruction of the gene. [1]

The dCas9 is asembled with the IPTG inducible promoter BBa_R0011, an RBS BBa_B0034 and a terminator BBa_B0010 making the expression of the protein controlable.

In 2019 the SDU-Denmark team used this IPTG inducible dCas9 protein in combination with a sgRNA to downregulate the expression og GFP and RFP.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 332
Illegal BglII site found at 1406
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3044009 short</partinfo>
-This dCas9 is a codon optimized version of BBa_K1150000 for expression in E. coli. Together with a single guide RNA (sgRNA), the protein can be used for silencing of specific target genes. dCas9 is the inactive form of the endonuclease Cas9. In contrast to Cas9, dCas9 does not cleave the DNA. Instead, it sterically blocks the elongation of transcription as a result of the sgRNA-DNA interaction. This will only repress the expression of the target gene thus avoiding the destruction of the gene [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4343198/].
+This dCas9 is a codon optimized version of <partinfo>BBa_K1150000</partinfo> for expression in E. coli. Together with a single guide RNA (sgRNA), the protein can be used for silencing of specific target genes. dCas9 is the inactive form of the endonuclease Cas9. In contrast to Cas9, dCas9 does not cleave the DNA. Instead, it sterically blocks the elongation of transcription as a result of the sgRNA-DNA interaction. This will only repress the expression of the target gene thus avoiding the destruction of the gene. [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4343198/]
-The dCas9 is asembled with the IPTG inducible promoter <partinfo>BBa_R0011</partinfo> and an RBS <partinfo>BBa_B0034</partinfo> making the expression of the protein controlable.
+The dCas9 is asembled with the IPTG inducible promoter <partinfo>BBa_R0011</partinfo>, an RBS <partinfo>BBa_B0034</partinfo> and a terminator <partinfo>BBa_B0010</partinfo> making the expression of the protein controlable.
+In 2019 the SDU-Denmark team used this IPTG inducible dCas9 protein in combination with a sgRNA to downregulate the expression og GFP and RFP.