Difference between revisions of "Part:BBa K1062002:Design"
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'''Figure 2: An example of a PAM sequence before the 20nt target sequence.''' | '''Figure 2: An example of a PAM sequence before the 20nt target sequence.''' | ||
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===References=== | ===References=== | ||
Revision as of 20:40, 17 September 2013
Guide RNA (gRNA) target for XylE (middle of gene)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.
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.
Figure 2: An example of a PAM sequence before the 20nt target sequence.