CRISPR/Cas gene editing is a relatively new technique which offers high accuracy in gene targeting and better target sequence selection, with less complexity and less off-target toxicity than previous genome editing technologies.
Highly targeted gene editing is promised with Amsbio’s Crispr gRNA lentivector cloning kits
Amsbio provides a standalone premade Cas9 expression lentivirus, and gRNA lentivector cloning kits, from which users can construct desired target gRNA lentivectors, to facilitate Crispr genomic editing.
These are the Amsbio Crispr gRNA lentivector cloning kits.
Application of both Cas9 lentivirus and gRNA lentivirus allows a target knockout or modification (knockin) when a donor DNA is also applied, the company says
These new kits are designed for rapid directional cloning of DNA duplex-encoded gRNA structure at efficiencies of greater than 95%.
The kit should eliminate the tedious preparation of a lentivector backbone, it adds.
The lentivector encodes an antibiotic marker or a dual marker (a fluorescent-antibiotic fusion marker) allowing generation of stable cell lines for long-term gRNA expression.
Two promoters are available – U6, and an inducible H1 promoter.
The 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.