Methods and kits for synthesis of siRNA expression cassettes

摘要

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.

主权项

1. A kit for a PCR-based approach for producing at least one promoter-containing siRNA expression cassette, comprising a double-stranded, promoter-containing template, a first oligonucleotide primer and a second oligonucleotide primer, wherein the double-stranded, promoter template has a sense strand and an antisense strand, wherein each of the sense strand and antisense strand has a 5′ end and a 3′ end, wherein the first oligonucleotide primer is complementary to the 3′ end of the antisense strand of the double-stranded, promoter-containing template, wherein the second oligonucleotide primer is complementary to the 3′ end of the antisense strand of the double-stranded, promoter-containing template, and wherein the second oligonucleotide primer comprises one or more nucleotide sequences complementary to a nucleotide sequence that encodes (1) a sense or antisense sequence of a double-stranded siRNA molecule and (2) a terminator sequence.