| abstract |
Recombinant constructs, vectors and expression cassettes are provided herein. The first comprises a first promoter (suitably a tRNA promoter operably linked to a first polynucleotide encoding a first single guide RNA) and a second encoding a Cas9 polypeptide. A second promoter is operably linked to the polynucleotide. The first single guide RNA includes a first portion that is complementary to the strand of the DNA virus target sequence and a second portion that can interact with the Cas9 polypeptide. Also provided are codon-optimized Cas9 polynucleotides and polypeptides from Staphylococcus aureus, which have a nuclear localization signal and optionally an epitope tag. Constructs for the production of sgRNA containing tRNA are also provided. Also provided are methods for inhibiting viral replication, inhibiting viral target sequence expression, or treating viral infections or virus-induced cancers using the compositions. |