CMV-CAS9D10A-2A-RFP Plasmid
SIGMA/CAS9D10ARFPP
Synonym: CAS9D10A Plasmid
Product Type: Chemical
Vector Map Features
| concentration | 20 ng/μL in TE buffer; DNA (1μg of plasmid DNA) |
| packaging | vial of 50 μL |
| promoter | Promoter name: CMV |
| Quality Level | 200  |
| recombinant | expressed in E. coli |
| reporter gene | RFP |
| selection | kanamycin |
| shipped in | dry ice |
| storage temp. | −20°C |
| Application: | Functional Genomics Use of Paired Cas9 Nickase + RFP for: • Creation of gene knockouts in multiple cell lines • Complete knockout of genes not amenable to RNAi • Creation of knock-in cell lines with promoters, fusion tags or reporters integrated into endogenous genes |
| General description: | The Cas9-D10A nickase plasmid co-expressing RFP uses the CMV promoter for strong transient expression of Cas9-D10A. Alternate promoters can be substituted by replacement of CMV using MluI and NheI. Also, the Cas9D10A expression plasmids can be linearized using XbaI for T7-based mRNA production. |
| Legal Information: | CRISPR Use License Agreement |
| Other Notes: | 1 vial containing 1ug of Cas9-D10A Nickase-RFP plasmid. Please note, this product does not contain any guide RNA sequence. A pair of gRNA plasmids must be purchased separately through the Custom CRISPR paired nickase product tab. |
| Other Notes: | The type II CRISPR/Cas system from the bacterium Streptococcus pyogenes has been engineered to function in eukaryotic systems using two molecular components: a single Cas9 protein and a non-coding guide RNA (gRNA). The Cas9 endonuclease can be programmed with a single gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB. RFP is co-expressed from the same mRNA as the Cas9 nickase protein via a 2A peptide linkage, enabling tracking of transfection efficiency and enrichment of genome editing activity in cell populations via fluorescence activated cell sorting (FACS). |
| Physical form: | 2 ug of Sigma Cas9-D10A nickase-RFP plasmid |
| RIDADR | NONH for all modes of transport |
| WGK Germany | nwg |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 41106609 |