Part:BBa_K1062000:Design
Guide RNA (gRNA) target for RFP
- 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_K1062000 is a gRNA containing a CSY4 cut site and specifically targets the RFP gene.
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.
The gRNA contains a CSY4 cut site that is essential to create the gRNA. Including in the part is a 20 nucleotide sequence that is targets specifically the RFP gene itself.
Sources
The RFP gRNA was retrieved from Stanley Qi's Lab at UCSF.
References
Currently Stanley Qi's lab is doing research in which hopes to use the CRISPR system in new applications.
[http://www.ncbi.nlm.nih.gov/pubmed/23452860 Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression ]
In the paper in the link provided, Stanley had shown to control gene expression using the CRISPR system using GFP and RFP as proof of concept. The RFP gRNA being used in our project is the same gRNA Stanley used in this paper.