Segmented micro RNA mimetics
This invention relates generally to segmented oligonucleotides capable of modulating gene expression. Specifically, the instant invention relates to segmented microRNA (miRNA) oligonucleotides, including segmented miRNA precursors and segmented pre-microRNAs. The invention also relates to compositions comprising such segmented oligonucleotides, as well as to methods of making and using such oligonucleotides for diagnosis and treatment of diseases associated or causally linked to aberrant levels or activities of gene expression, including aberrant levels of coding and/or non-coding RNA.
1. A double-stranded oligonucleotide molecule represented by Formula III:
wherein
(a) the molecule comprises a contiguous passenger strand and a guide strand where each line and its adjacent “N” represent a contiguous stretch of nucleotides, each of “X1,” “X2” and “X3” represent the number of nucleotides in each stretch, “GIN” represents a discontinuity in the guide strand, “Y1” represents a number of nucleotide positions in the discontinuity, and each group of dashed lines “ ” and its adjacent “(W)” represents a terminal overhang that is optionally present, and each of “Z1” and “Z2” represents the number of overhanging nucleotides;
(b) X1 is an integer from 16 to 26, X2 is an integer from 2 to 21, X3 is an integer from 2 to 9, Y1 is an integer from 0 to 6, provided that the sum of X2, X3 and Y1 is an integer from 16 to 26;
Z1 and Z2 are independently integers from 0 to 4;
(c) N X2 and N X3 together comprise sequence having at least 6 contiguous nucleotides of the seed sequence of a miRNA sequence having any of SEQ ID NOs: 1-1090;
(d) N X2 and N X3 together comprise sequence having at least 50% homology to the miRNA sequence; and
(e) N X1 comprises sequence having at least 50% complementarity to the miRNA sequence.
2. The double stranded oligonucleotide molecule of claim 1 , wherein G/N comprises a nick in the guide strand.
3. The double stranded oligonucleotide molecule of claim 1 , wherein G/N comprises one or more gap(s) in the guide strand.
4. The double stranded oligonucleotide molecule of claim 1 , wherein G/N comprises one or more nonnucleotide substitutions in the guide strand.
5. The double stranded oligonucleotide molecule of claim 1 , wherein G/N comprises one or more nonnucleotide insertions in the guide strand.
6. The double stranded oligonucleotide molecule of claim 1 , wherein NX2 and NX3 together comprise a sequence having at least 7 contiguous nucleotides of the seed sequence of the miRNA.
7. The double stranded oligonucleotide molecule of claim 1 , wherein NX2 and NX3 together comprise a sequence having at least 8 contiguous nucleotides of the seed sequence of the miRNA.
8. The double stranded oligonucleotide molecule of claim 1 , wherein NX2 and NX3 together comprise a sequence having at least 60, 70, 80, or 90% homology to the miRNA sequence.
9. The double stranded oligonucleotide molecule of claim 1 , wherein NX 1 comprises a sequence having at least 60, 70, 80, or 90% complementarity to the miRNA sequence.
10. A composition comprising the double stranded oligonucleotide molecule of claim 1 and a pharmaceutically acceptable carrier, diluent, excipient, adjuvant, emulsifier, buffer, stabilizer, or preservative.
11. The double stranded oligonucleotide molecule of claim 5 , wherein the one or more nonnucleotide insertions in the guide strand comprise an alkyl moiety.
12. The double stranded oligonucleotide molecule of claim 11 , wherein the alkyl moiety comprises any of C 1 -C 20 .
13. The double stranded oligonucleotide molecule of claim 11 , wherein the alkyl moiety comprises C 3 , C 4 , C 5 , or C 6 .
14. A double-stranded oligonucleotide molecule represented by Formula III:
wherein
(a) the molecule comprises a contiguous passenger strand and a guide strand where each line and its adjacent “N” represent a contiguous stretch of nucleotides, each of “X1,” “X2” and “X3” represent the number of nucleotides in each stretch, “GIN” represents a discontinuity in the guide strand, “Y1” represents a number of nucleotide positions in the discontinuity, and each group of dashed lines “ ” and its adjacent “(W)” represents a terminal overhang that is optionally present, and each of “Z1” and “Z2” represents the number of overhanging nucleotides;
(b) X 1 is an integer from 16 to 26, X2 is an integer from 2 to 21, X3 is an integer from 2 to 9, Y1 is 0, 1, or 2, provided that the sum of X2, X3 and Y1 is an integer from 16 to 26; Z1 and Z2 are independently integers from 0 to 4;
(c) NX2 and NX3 together comprise sequence having at least 6 contiguous nucleotides of the seed sequence of a miRNA sequence having any of SEQ ID NOs: 1-1090;
(d) NX2 and NX3 together comprise sequence having at least 50% homology to the miRNA sequence;
(e) NX1 comprises sequence having at least 50% complementarity to the miRNA sequence; and
(f) G/N comprises an alkyl moiety.
15. The double stranded oligonucleotide molecule of claim 14 , wherein the alkyl moiety comprises any of C 1 -C 20 .
16. The double stranded oligonucleotide molecule of claim 14 , wherein the alkyl moiety comprises C 3 , C 4 , C 5 , or C 6 .
17. The double stranded oligonucleotide molecule of claim 1 , wherein NX3 comprises sequence having at least 6 contiguous nucleotides of the seed sequence.