CRISPR UNIVERSAL NEGATIVE CONTROL 1
SIGMA/CRISPR06
Product Type: Chemical
Vector Map Features
| application(s) | CRISPR |
| concentration | 20 ng/μL in TE buffer; DNA (1μg of plasmid DNA) |
| form | liquid |
| packaging | vial of 50 μL |
| Quality Level | 200  |
| shipped in | dry ice |
| storage temp. | −20°C |
| Application: | Functional Genomics/Target Validation • Creation of gene knockouts in cell lines • Creation of knock-in cell lines with promoters, fusion tags or reporters integrated into endogenous genes |
| Biochem/physiol Actions: | CRISPR/Cas systems are employed by bacteria and archaea as a defense against invading viruses and plasmids. Recently, 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. |
| Features and Benefits: | Allows for expression of Cas9 and GFP without specific targeting of the CRISPR/Cas9 system. |
| General description: | Universal negative control CRISPRs have been designed not to recognize any sequence in the human, mouse or rat genome. A single vector format is provided which includes the Cas9 expression cassette and gRNA sequence. This vector includes GFP which is co-expressed from the same mRNA as the Cas9 protein via a 2A peptide linkage and enables for tracking of transfection efficiency or enrichment in cell populations via FACS. |
| Legal Information: | CRISPR Use License Agreement Lentiviral and WPRE License Agreements |
| Other Notes: | 1 vial containing 1ug of U6-gRNA/CMV-Cas9-GFP plasmid expressing a non-targeting guide sequence. |
| Other Notes: | Typical transfection concentrations used in literature are in the ranges of 2.0 to 8.0ug of the single vector expressing the guideRNA and Cas9 . (All dosages above assume 0.5 to 1 million cells nucleofected) |
| Physical form: | Sigma U6-gRNA/CMV-Cas9-GFP plasmid expressing a non-targeting guide sequence supplied at a concentration of 20ng/ul in 50ul. |
| Preparation Note: | Sigma CRISPR plasmid products are delivered as mini-prep aliquots, which may not be suitable for transfection into particular cell types. For best results, we advise maxi-prepping plasmids using endotoxin-free DNA purification kits prior to transfection. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 41106609 |