RNA
XylE ib

Part:BBa_K1062003:Design

Designed by: David Dinh   Group: iGEM13_UCSF   (2013-09-17)
Revision as of 20:41, 17 September 2013 by Dadinh (Talk | contribs) (→‎Design Notes)

Guide RNA (gRNA) target for XylE (end of gene)


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

Part:BBa_K1062002 is a gRNA that specifically targets the middle of the XylE gene.

gRNA

A gRNA is a 102 nt sequence in which contains: a 20 nt sequence which will contain the sequence of the intended target, a 42 nt Cas9-binding hairping, and a 40 nt transcription terminator.

GRNA with NNN sequence.png

Figure 1: An image show a general picture of a gRNA.


An important thing to note when using or creating a gRNA is that before the 20nt sequence, there must be a 3nt sequence called a PAM sequence which contain this sequence 5'-NGG-3'. This 3nt sequence is what the Cas9 use as a binding signal. It is also important to point out that the PAM sequence is never in the gRNA itself.

Pam.png

Figure 2: An example of a PAM sequence before the 20nt target sequence.

Source

baah

References