CRISPR07

CRISPR UNIVERSAL NEGATIVE CONTROL 2

• UNSPSC kód: 41106609
• NACRES: NA.51

Tulajdonságok

• form: liquid
• Minőségi szint: 200
• kiszerelés: vial of 50 μL
• koncentráció: 20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
• alkalmazás(ok): CRISPR
• kiszállítva: dry ice
• tárolási hőmérséklet: −20°C

Leírás

Általános leírás

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.

Alkalmazás

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

Tulajdonságok és előnyök

Allows for expression of Cas9 and GFP without specific targeting of the CRISPR/Cas9 system.

Komponensek

1 vial containing 1ug of U6-gRNA/CMV-Cas9-GFP plasmid expressing a non-targeting guide sequence.

Elv

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.

Fizikai forma

Sigma U6-gRNA/CMV-Cas9-GFP plasmid expressing a non-targeting guide sequence supplied at a concentration of 20ng/ul in 50ul.

Elkészítési megjegyzés

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.

Egyéb megjegyzések

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)

Jogi információk

CRISPR Use License Agreement

Lentiviral and WPRE License Agreements

Biztonsági adatok

• Tárolási osztály kódja: 12 - Non Combustible Liquids
• WGK: WGK 1
• Lobbanási pont (F): Not applicable
• Lobbanási pont (C): Not applicable