DNA

Part:BBa_K4800013

Designed by: Wenbin Zhao   Group: iGEM23_NJTech-China-B   (2023-10-06)
Revision as of 09:55, 12 October 2023 by ZQM 0907 (Talk | contribs)


YcjQ sgRNA

SgRNA, which is small guide RNA, is an important component of the CRISPR knockdown system. The N20 sequence of the YcjQ SgRNA is derived from the YcjQ gene and can be localised to the YcjQ gene in the Escherichia coli NT1003 genome to assist the Cas9 protein in silencing the gene.


Document

YcjQ was involved in the 1,5-PDO degradation. In order to increase the yield, we decided to knock out the YcjQ gene. We decided to use CRISPR/Cas9 to construct the strain E. coli NT1003 ΔYcjQ.

Build:

Fig.1 YcjQ sgRNA

We designed the N20 sequence of YcjQ by using the CHOPCHOP website (http://chopchop.cbu.uib.no), synthesized them on primers and ligated the N20 into the pTarget-sgRNA plasmid by means of PCR.

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]


[edit]
Categories
Parameters
None