Abstract: This invention relates to interfering RNA (iRNA) molecules and their applications, especially multi-targets iRNA molecules and their applications. The said multi-targets iRNA molecules comprised of a sense strand annealed onto at least one antisense strand, each strand is at least 30 nucleotides in length, the sense or antisense strand has at least two segments, which can target at least two RNAs of different genes, or can target at least two portions of an RNA, and wherein the iRNA does not induce an interferon-response when transfected into a cell. The iRNA molecule can interfere with the translation procedure post-transcription, and the target gene is inhibited or blocked, the iRNA does not induce an interferon-response in vivo. The RNA molecules are the active ingredient in preparation of the drug which can regulate one or many genes function.

Abstract: The present invention provides a PCR based high-throughput method for preparing full-sites siRNA polynucleotide pool, comprising: DNase I random digestion; Loop-1 phosphate linker ligation; single PCR amplification; a type III restriction/modification enzyme digestion; blunt ending; Loop-2 phosphate linker ligation; double primer PCR; FokI digestion and cloning into an siRNA expression vector. The present invention enables the use of a type III restriction/modification enzyme linkers mediated PCR method for high-throughput preparing an siRINA polynucleotide pool, in which the functional length of siRNAs can be controllably distributed from 19-23 bp, thus completely mimic the natural siRNA length diversity, specially suitable for RNAi therapeutic targets screening. The present invention overcomes the bottlenecks and drawbacks of conventional siRNA polynucleotide pool construction technologies.

Abstract: This invention relates to interfering RNA (iRNA) molecules and their applications, especially multi-targets iRNA molecules and their applications. The said multi-targets iRNA molecules comprised of a sense strand annealed onto at least one antisense strand, each strand is at least 30 nucleotides in length, the sense or antisense strand has at least two segments, which can target at least two RNAs of different genes, or can target at least two portions of an RNA, and wherein the iRNA does not induce an interferon-response when transfected into a cell. The iRNA molecule can interfere with the translation procedure post-transcription, and the target gene is inhibited or blocked, the iRNA does not induce an interferon-response in vivo. The RNA molecules are the active ingredient in preparation of the drug which can regulate one or many genes function.

Abstract: The present invention provides a PCR based high-throughput method for preparing full-sites siRNA polynucleotide pool, comprising: DNase I random digestion; Loop-1 phosphate linker ligation; single PCR amplification; a type III restriction/modification enzyme digestion; blunt ending; Loop-2 phosphate linker ligation; double primer PCR; FokI digestion and cloning into an siRNA expression vector. The present invention enables the use of a type III restriction/modification enzyme linkers mediated PCR method for high-throughput preparing an siRINA polynucleotide pool, in which the functional length of siRNAs can be controllably distributed from 19-23 bp, thus completely mimic the natural siRNA length diversity, specially suitable for RNAi therapeutic targets screening. The present invention overcomes the bottlenecks and drawbacks of conventional siRNA polynucleotide pool construction technologies.

Abstract: This invention relates to the application of the highly conserved sequences of viral genome, especially from a highly conserved domain of enteroviral genome as templates to design target small ligand RNAs (sliRNAs). The resulting sliRNAs are therapeutically active ingredients in the treatment of the related diseases caused by pathological angiogenesis.

Abstract: The present invention provides a PCR based high-throughput method for preparing full-sites siRNA polynucleotide pool, comprising: DNase I random digestion; Loop-1 phosphate linker ligation; single PCR amplification; a type III restriction/modification enzyme digestion; blunt ending; Loop-2 phosphate linker ligation; double primer PCR; FokI digestion and cloning into an siRNA expression vector. The present invention enables the use of a type III restriction/modification enzyme linkers mediated PCR method for high-throughput preparing an siRNA polynucleotide pool, in which the functional length of siRNAs can be controllably distributed from 19-23 bp, thus completely mimic the natural siRNA length diversity, specially suitable for RNAi therapeutic targets screening. The present invention overcomes the bottlenecks and drawbacks of conventional siRNA polynucleotide pool construction technologies.

Abstract: This invention provides novel methods for producing a plurality of polynucleotides prepared from a polynucleotide sample and the plurality of polynucleotides so produced. In one embodiment, the plurality of polynucleotides is prepared by subtractive hybridization between test and reference polynucleotide samples and is substantially enriched in sequences that are either not present in the reference polynucleotide sample or are present in the reference polynucleotide sample in substantially lower concentration than in the test polynucleotide sample. The plurality of polynucleotides is also substantially enriched in low-abundance sequences, relative to the test polynucleotide sample. The invention also provides kits useful in the methods of the invention and for using the polynucleotides produced thereby. The polynucleotides are useful in a wide variety of applications, such as cloning, expression, and hybridization studies.

Abstract: This invention provides novel methods for producing a plurality of polynucleotides prepared from a polynucleotide sample and the plurality of polynucleotides so produced. The plurality of polynucleotides is either substantially enriched in high-abundance polynucleotide sequences relative to the polynucleotide sample or substantially enriched in low-abundance polynucleotide sequences relative to the polynucleotide sample. Each of the polynucleotides in the plurality of polynucleotides includes a RNA promoter sequence and a universal primer site. The invention also provides kits useful in the methods of the invention and for using the pools of polynucleotides produced thereby. The polynucleotide pools are useful in a wide variety of applications, such as cloning, expression, and hybridization studies. Pools enriched in low-abundance polynucleotides are particularly useful in studies aimed at identifying important regulatory proteins.