Difference between revisions of "Part:BBa K2361000"

Revision as of 08:54, 30 October 2017

spdCas9

This part contains the protein coding sequence for the dCas9 originating from Streptococcus pyogenes. This protein is a catalytically-dead Cas9 variant, which lacks endonuclease activity. It is used for gene repression using CRISPR-interference.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1100
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 3379
Illegal XhoI site found at 4115
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2361000 short</partinfo>
-This part contains the protein coding sequence for the dCas9 originating from Streptococcus pyogenes. This protein is a catalytically-dead Cas9 viriant. This viriant lacks endonuclease activity and therefor is not able to cut the DNA upon binding through association with a single guide RNA [sgRNA] and its complementary sequence.
+This part contains the protein coding sequence for the dCas9 originating from <i>Streptococcus pyogenes</i>. This protein is a catalytically-dead Cas9 variant, which lacks endonuclease activity. It is used for gene repression using CRISPR-interference.
 <!-- Add more about the biology of this part here
 ===Usage and Biology===
+Overexpression of dCas9 may cause cytotoxic effects [1], therefore we recommend expressing it using an inducible promotor.
+In this part the start codon (ATG) starts directly behind the XbaI site and not at the last A of the XbaI site. This may lead to reduced efficiency of expression when combined with RBS's from the iGEM repository, but more importantly this needs to be taken into consideration when creating fusion proteins with this part to ensure that no frame-shifts occur.
+[1] Nielsen, A. A., & Voigt, C. A. (2014). Multi-input CRISPR/Cas genetic circuits that interface host regulatory networks. Molecular Systems Biology, 10(11), 763. http://doi.org/10.15252/msb.20145735
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