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Abingdon, 2016 – Anglo-American life science firm AMSBIO has introduced new CRISPR gRNA Lentivector Cloning Kits that provide scientists with a targeted and precise genomic gene editing methodology.
Targeted and precise genomic gene editing technologies are today key tools for genomic correction, modification and gene therapy. The recently discovered CRISPR/Cas gene editing technique offers higher targeting accuracy, much more target sequence selection, much less complexity and less off-target cell toxicity than the previous generation genome editing technologies.
To enable CRISPR genomic editing, the AMSBIO kit provides the standalone premade Cas9 expression lentivirus, and the gRNA lentivector cloning kits from which you can construct your desired target gRNA lentivectors. Applying both Cas9 lentivirus and gRNA lentivirus allows you to achieve the target knockOut or modification (knockIn) when a donor DNA is also applied.
The new kits enable rapid, precisely directional cloning of your DNA duplex encoded gRNA structure at efficiencies of greater than 95%. Also incorporating a pre-linearized lentivector the kit eliminates the need for tedious preparation of a lentivector backbone. The lentivector included in each AMSBIO kit encodes an antibiotic marker or a dual marker (a fluorescent-antibiotic fusion marker) allowing generation of stable cell lines for long-term gRNA expression. There are two promoters available: U6 and an optional inducible H1 promoter. The gRNA lentivectors driven by the inducible H1 promoter can be used for constitutive high expression without the need for induction. This modified H1 promoter allows the inducible expression of gRNA when the tetracycline repressor protein (TetR) is present in advance and the U6 promoter is the constitutive promoter.