| 摘要 |
The present disclosure provides a non-naturally occurring miRNA having a stem-loop structure comprising a scaffold derived from a first endogenous miRNA (e.g., miR-196a-2 or miR-204), a mature strand derived from a second endogenous miRNA, and a star strand sequence that is at least partially complementary to the mature strand sequence. The present disclosure also provides a non-naturally occurring miRNA having a stem-loop structure comprising a scaffold derived from an endogenous miRNA (e.g., miR-196a-2 or miR-204), a mature strand designed to be at least partially complementary to a target RNA, and a star strand sequence that is at least partially complementary to the mature strand sequence. The methods and compositions of the disclosure may be used to mediate gene silencing via the RNAi pathway. |
| 主权项 |
1. A method of delivery of a mature endogenous miRNA to a cell, the method comprising:
introducing in said cell a non-naturally occurring miR-196a-2 miRNA, wherein said non-naturally occurring miR-196a-2 miRNA comprises a nucleic acid having a stem-loop structure, wherein the stem of the stem-loop structure incorporates a miRNA-196-a-2 scaffold, and a mature strand-star strand duplex, wherein said mature strand has a sequence derived from a mature endogenous miRNA, wherein said mature endogenous miRNA has a sequence that is distinct from the sequence of the endogenous mature strand of miR-196a-2, wherein position 1 of the mature strand is U and the opposite nucleotide in the star strand is G and the duplex contains a mismatch at position 12 of the mature strand and either no additional mismatches or wobble pairs at any position other than positions 1 and 12 of the mature strand, or mismatches or wobble pairs at one or more of positions other than at positions 2, 3, 4, 6, 7, 8, 9, 10, 11, 13, 14, 15, 16 and 17 within the duplex, wherein positions of the mature strand are numbered relative to the 5′ end of the mature strand, and the mature strand is about 19-25 nucleotides in length; wherein said non-naturally occurring miR-196a-2 miRNA is processed in said cell to yield a mature miRNA having the sequence of said mature strand. |
