Nucleic acids for down-regulation of gene expression
AssigneesUniversity of Pretoria, Universite De Geneve
InventorsKarl-Heinz Krause, Renier Myburgh, Patrick Salmon
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Abstract
Recombinant nucleic acid molecules are provided that form hair pin structures and can be used to down-regulate gene expression. For example, a nucleic acid molecule can comprise a flanking and lower stem loop sequence from a mir-16 gene; an antisense target sequence; a mir-30 loop sequence; a complement of the anti-sense target sequence; and a lower stem loop complementary to the mir-16 sequence. Methods for down regulating gene expression in a cell using such recombinant nucleic acid molecules are also provided.
Claims60
A recombinant nucleic acid molecule comprising from 5′ to 3′ and in the order from (a)-(g):
- (a) a mir-16 flanking sequence comprising the sequence of SEQ ID NO:
- 1;
- (b) a first lower stem sequence comprising the mir-16 sequence of SEQ ID NO:
- 2, SEQ ID NO:
- 3, SEQ ID NO:
- 4, SEQ ID NO:
- 5 or SEQ ID NO:
- 6;
- (c) an anti-sense target sequence 22 nucleotides in length;
- (d) a mir-30 loop sequence comprising the sequence of SEQ ID NO:
- 7;
- (e) a sense sequence wherein the sequence is complementary to the sequence of (c) except that the sequence comprises one or two mismatches relative to the sequence of (c), wherein the one or two mismatches comprise:
- i) a mismatch located at the position 8 to 14 of the sense sequence;
- or
- ii) a mismatch at the final 3′ position (position 22) of the sense sequence;
- (f) a second lower stem sequence wherein the sequence is complementary to the sequence of (b);
- and
- (g) a second flanking sequence.
A recombinant nucleic acid molecule comprising from 5′ to 3′ and in the order from (a)-(g):
- (a) a mir-16 flanking sequence;
- (b) a first lower stem sequence comprising a mir-16 sequence;
- (c) an anti-sense target sequence 22 nucleotides in length;
- (d) a mir-30 loop sequence;
- (e) a sense sequence wherein the sequence is complementary to the sequence of (c) except that the sequence comprises one or two mismatches relative to the sequence of (c), wherein the one or two mismatches comprise:
- i) a mismatch located at the position 8 to 14 of the sense sequence;
- or
- ii) a mismatch at the final 3′ position (position 22) of the sense sequence;
- (f) a second lower stem sequence wherein the sequence is complementary to the sequence of (b), wherein the lower stem is at least 11 nucleotides in length;
- and
- (g) a second flanking sequence.
Dependencies are inferred from the claim text.