Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Shihong Chen
Date created: 2013-07-10 11:00:00
Date modified: 2015-05-08 01:08:55
protein dCas9.Two mutations,D10A and H841A,from Cas9 gene without stop codon.
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K1051800_sequence (Version 1) |
Description
Protein dCas9 is the key of the technology CRISPRi,and this part is the gene of the dcas9.Delete its ending codon is to provide an convenient way to add other protein domain to improve the function of dCas9.Notes
To mutation four Restriction Enzyme cutting sites(SpeI) and two codons(D10A & H841A),we design PCR primers to mutate them by PCR using Phution DNA polymerase.Refrence http://www.ncbi.nlm.nih.gov/pubmed/?term=repurposing+CRISPR+as+an+RNA-Guided+Platfrom
Source
The Cas9 gene was got from Mr. Kai Tian,BGI Shenzhen China.The gene was splited into three parts to synthetize optimizing the codons in eucaryon.