CAS9GFPV

Cas9 GFP Lentiviral Particles

• Synonym(s): GFP Lentiviral Particles
• UNSPSC Code: 41106621
• NACRES: NA.51

Properties

• form: liquid
• packaging: pkg of 8 x 25 μL (aliquots)
• concentration: ≥5 x 10 ^{8 VP/ml virus (via p24 assay)}
• application(s): CRISPR
• reporter gene: GFP
• shipped in: dry ice
• storage temp.: −70°C

Description

General description

Ready-to-use Cas9-GFP lentiviral particles enable immediate transduction of a wide range of cell lines. Cas9-GFP lentiviral particles efficiently and stably integrate Cas9 and GFP linked by a 2A peptide and driven by the EF1 alpha promoter (EF1a-Cas9-2A-GFP) for strong expression in both dividing and non-dividing cell lines. They are ideal for avoiding the tedious lentivirus production process and are one part of a two part CRISPR system with individual Cas9 and gRNA expression vectors. For easy fluorescence microscopy a MOI greater than 200 should be employed.

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Application

Functional Genomics/Target Validation

• Creation of gene knockouts in multiple cell lines
• Complete knockout of genes not amenable to RNAi
• Creation of cell lines stably expressing Cas9-GFP

Features and Benefits

• Highly specific
• Highly active
• Ready to infect/transduce

Principle

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.

Unit Definition

VP/ml is the concentration unit of measure for viral titer estimated by p24 assay.

Legal Information

CRISPR Use License Agreement

Evrogen Fluorophore Licenses

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 3
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable