RNA

Part:BBa_K1994013:Design

Designed by: Liam Carroll   Group: iGEM16_Warwick   (2016-09-22)


sgRNA with dCas9 binding site sequence 9 and PP7 handle insert


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]


Design Notes

To simplify the traditional gRNA comprised of a crRNA containing a dCas9 handle, and tracrRNA which binds to the PAM sequence in CRISPR, we created a joined sgRNA. As modifying the 3' end of the gRNA does not affect dCas9 binding we have used a deactivated dcas9 terminator and placed golden gate adaptable sites. The two sites are located where the complementary PAM-proximal sequence is located, allowing the distance dCas9 binds from the promoter to be adapted.The other golden gate adaptor in the position after the deactivated terminator so any RNA sequence such as an RNA binding protein handle can be inserted. This region is then followed by a real terminator for the strand.

Source

Completely synthetic

References