METHODS AND KITS FOR SYNTHESIS OF siRNA EXPRESSION CASSETTES
AssigneeCity of Hope
InventorsJohn J. ROSSI, Daniela CASTANOTTO
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Abstract
Amplification-based methods and kits for rapidly producing siRNA expression cassettes are provided. Also provided are methods for expressing amplified siRNA expression cassettes in cells.
Claims6
A PCR-based approach in the form of a kit for producing a promoter-containing siRNA expression cassette, comprising a double-stranded, promoter-containing template, an oligonucleotide primer complementary to the 5′ end of the promoter-containing template, and an oligonucleotide primer complementary to the 3′ end of the promoter-containing template, wherein the 3′ primer comprises one or more sequences complementary to a sequence encoding a sense and/or antisense sequence of a siRNA or siRNA molecule.
A method for screening potential target sequences susceptible to siRNA mediated degradation, comprising transfecting a cell with an amplified siRNA expression cassette under conditions in which an siRNA molecule can be expressed and mediate degradation of the potential target sequences.
A method for inhibiting expression of a target gene, comprising transfecting a cell with an amplified siRNA expression cassette under conditions in which an siRNA molecule can be expressed and inhibit expression of the target gene.
Dependencies are inferred from the claim text.