Abstract: Methods for the production of duplexes and single-stranded RNA and/or DNA of a desired length and sequence based on a novel template design which incorporates 2 polymerase promoters, primers, and production sequences within a single molecule are provided. This Single-stranded Template molecule design allows high-efficiency, high-yield production of single or multiple nucleic acid molecules in a single reaction vessel and thus is amenable to high-throughput automation. This Single-stranded Template molecule design also allows easy incorporation of Single-stranded Template molecules into delivery vectors for either in vitro, ex vivo, in vivo, or therapeutic application. Methods for producing Single stranded Template molecule-based RNA or DNA molecules, or hybrid molecules, in vivo and therapeutic uses for such molecules are provided. Single-stranded Template molecule kit designs are also described.

Abstract: A method is provided for the induction of aberrant RNA interference (abRNAi) by introducing ‘aberrant’ RNAs into the cell wherein a long-stranded antisense or sense RNA is simultaneously introduced with a short 4-12 mer homologous or complementary or random RNA wherein the long-short RNAs induce sequence specific inhibition of the homologous gene within a cell. The invention disclosed herein can be used to suppress gene expression in vitro, ex vivo or in vivo.

Abstract: Methods for the production of duplexes and single-stranded RNA and/or DNA of a desired length and sequence based on a novel template design which incorporates 2 polymerase promoters, primers, and production sequences within a single molecule are provided. This single molecule template design allows high-efficiency, high-yield production of single or multiple nucleic acid molecules in a single reaction vessel and thus is amenable to high-throughput automation. This single molecule design also allows easy incorporation of single molecule templates into delivery vectors for either in vitro, ex vivo, in vivo, or therapeutic application. Methods for producing single template molecule-based RNA or DNA molecules, or hybrid molecules, in vivo and therapeutic uses for such molecules are provided. Single molecule template kit designs are also described.