Methods and compositions for mediating gene silencing
AssigneeUniversity of Massachusetts
InventorTariq Rana
Life of this patent
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Abstract
The present invention provides methods of conducting RNAi using siRNAs that are sequentially administered as single-stranded oligonucleotides. The siRNAs can be canonical or have non-canonical ends. The compositions and methods of the invention can bypass activation of interferon pathways and yet still efficiently and specifically activate RNAi/gene silencing. In another embodiment, the siRNAs of the invention are modified to allow for the calculation of certain RNAi activities, e.g., RISC activity. The invention also provides methods of using the compositions in research, diagnostic, and therapeutic applications.
Claims40
A small interfering RNA (siRNA), comprising a sense strand and an antisense strand, the antisense strand having a sequence sufficiently complementary to a target gene sequence to direct target-specific RNA interference (RNAi), wherein the strands, when aligned, form at least one non-canonical end.
A small interfering RNA (siRNA), comprising a sense strand and an antisense strand, the antisense strand having a sequence sufficiently complementary to a target gene sequence to direct target-specific RNA interference (RNAi), wherein the sense strand and antisense strand are separately and temporally exposed to the target gene sequence.
A composition comprising the siRNA molecule of any the preceding claims and a pharmaceutically acceptable carrier.
A method of activating target-specific RNA interference (RNAi) in a cell comprising,
- introducing into the cell a small interfering RNA (siRNA), comprising a sense strand and an antisense strand, the antisense strand having a sequence sufficiently complementary to a target gene sequence to direct target-specific RNA interference (RNAi), wherein the strands, when aligned, form at least one non-canonical end,
- the siRNA being introduced in an amount sufficient for degradation of target mRNA to occur, thereby activating target-specific RNAi in the cell.
A method of validating a candidate protein as a suitable target for drug discovery comprising,
- introducing into a cell or organism the siRNA of any one of the preceding claims, and
- maintaining the cell or organism under conditions such that target-specific RNAi can occur,
- determining a characteristic or property of the cell or organism, and
- comparing the characteristic or property to a suitable control,
- the comparison yielding information about whether the candidate protein is a suitable target for drug discovery.
A kit comprising reagents for activating target-specific RNA interference (RNAi) in a cell or organism, the kit comprising:
- the siRNA molecule of any one of the preceding claims, and
- instructions for use.
Dependencies are inferred from the claim text.