Novel polynucleotides, cells and methods
AssigneeLaverock Therapeutics Limited
InventorsTimothy Allsopp, Vlad Seitan, Vishal Menon, Anna Klucnika
Life of this patent
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Abstract
There are disclosed inter alia novel inhibitory RNAs, or a polynucleotide sequence encoding an inhibitory RNA, and expression cassettes, vectors, virions, synthetic microRNA, synthetic pre-microRNA molecules and cells, and associated medical uses and methods of modifying protein expression, especially B2M protein expression.
Claims69
An inhibitory RNA, or a polynucleotide sequence encoding an inhibitory RNA, preferably a microRNA, the inhibitory RNA comprising or consisting a sequence selected from:
- (i) any one of SEQ ID Nos:
- 1-17, 64-127;
- (ii) a variant of any one of SEQ ID Nos:
- 1-17, 64-127;
- (iii) a fragment of any one of SEQ ID Nos:
- 1-17, 64-127;
- and
- (iv) a DNA sequence complementary to any one of (i), (ii) or (iii).
The inhibitory RNA or a polynucleotide sequence encoding an inhibitory RNA according to any preceding claim, which is a synthetic nucleic acid sequence.
The inhibitory RNA or polynucleotide sequence encoding an inhibitory RNA according to any preceding claim, which comprises or encodes a microRNA scaffold.
The inhibitory RNA or polynucleotide sequence encoding an inhibitory RNA according to any preceding claim, wherein the polynucleotide of any one of SEQ ID NOs:
- 1-17, 64-127 is inserted into a human pre-microRNA sequence, preferably wherein:
- (i) SEQ ID NO:
- 1 or 7 is inserted into hsa-mir-191 (SEQ ID NO:
- 18 or 24, respectively);
- (ii) SEQ ID NO:
- 2 or 11 is inserted into hsa-mir-302a (SEQ ID NO:
- 19 or 28, respectively);
- (iii) SEQ ID NO:
- 3 or 12 is inserted into hsa-mir-302c (SEQ ID NO:
- 20 or 29, respectively);
- (iv) SEQ ID NO:
- 4 or 16 is inserted into hsa-mir-93 (SEQ ID NO:
- 21 or 33, respectively);
- (v) SEQ ID NO:
- 5 is inserted into hsa-mir-106a (SEQ ID NO:
- 22);
- (vi) SEQ ID NO:
- 6 is inserted into hsa-mir-106b (SEQ ID NO:
- 23);
- (vii) SEQ ID NO:
- 8 is inserted into hsa-mir-200c (SEQ ID NO:
- 25);
- (viii) SEQ ID NO:
- 9 is inserted into hsa-mir-20a (SEQ ID NO:
- 26);
- (iv) SEQ ID NO:
- 10 is inserted into hsa-mir-21 (SEQ ID NO:
- 27);
- (x) SEQ ID NO:
- 13 is inserted into hsa-mir-363 (SEQ ID NO:
- 30);
- (xi) SEQ ID NO:
- 14 is inserted into hsa-mir-518b (SEQ ID NO:
- 31);
- (xii) SEQ ID NO:
- 15 is inserted into hsa-mir-744 (SEQ ID NO:
- 32);
- (xiii) SEQ ID NO:
- 17 is inserted into hsa-mir-99b (SEQ ID NO:
- 34);
- (xiv) SEQ ID NO:
- 64, SEQ ID NO:
- 73 or SEQ ID NO:
- 89 is inserted into hsa-mir-320a (SEQ ID NO:
- 128, SEQ ID NO:
- 137 or SEQ ID NO:
- 153 respectively);
- (xv) SEQ ID NO:
- 65, SEQ ID NO:
- 77 or SEQ ID NO:
- 94 is inserted into hsa-mir-520f (SEQ ID NO:
- 129, SEQ ID NO:
- 141 or SEQ ID NO:
- 158 respectively);
- (xvi) SEQ ID NO:
- 66, SEQ ID NO:
- 78 or SEQ ID NO:
- 95 is inserted into hsa-mir-652 (SEQ ID NO:
- 130, SEQ ID NO:
- 142 or SEQ ID NO:
- 159 respectively);
- (xvii) SEQ ID NO:
- 67 is inserted into hsa-mir-106a (SEQ ID NO:
- 131);
- (xviii) SEQ ID NO:
- 68 or SEQ ID NO:
- 79 is inserted into hsa-mir-1180 (SEQ ID NO:
- 132 or SEQ ID NO:
- 143 respectively);
- (xix) SEQ ID NO:
- 69 or SEQ ID NO:
- 81 is inserted into hsa-mir-15b (SEQ ID NO:
- 133 or SEQ ID NO:
- 145 respectively);
- (xx) SEQ ID NO:
- 70, SEQ ID NO:
- 82 or SEQ ID NO:
- 99 is inserted into hsa-mir-182 (SEQ ID NO:
- 134, SEQ ID NO:
- 146 or SEQ ID NO:
- 163 respectively);
- (xxi) SEQ ID NO:
- 71 or SEQ ID NO:
- 87 is inserted into hsa-mir-23a (SEQ ID NO:135 or SEQ ID NO:
- 151 respectively);
- (xxii) SEQ ID NO:
- 72 is inserted into hsa-mir-26b (SEQ ID NO:
- 136);
- (xxiii) SEQ ID NO:
- 74 or SEQ ID NO:
- 90 is inserted into hsa-mir-335 (SEQ ID NO:
- 138 or SEQ ID NO:
- 154 respectively);
- (xxiv) SEQ ID NO:
- 75 or SEQ ID NO:
- 91 is inserted into hsa-mir-361 (SEQ ID NO:
- 139 or SEQ ID NO:
- 155 respectively);
- (xxv) SEQ ID NO:
- 76 is inserted into hsa-mir-518b (SEQ ID NO:
- 140);
- (xxvi) SEQ ID NO:
- 80 is inserted into hsa-mir-1307 (SEQ ID NO:
- 144);
- (xxvii) SEQ ID NO:
- 83 is inserted into hsa-mir-205 (SEQ ID NO:
- 147);
- (xxviii) SEQ ID NO:
- 84 is inserted into hsa-mir-22 (SEQ ID NO:
- 148);
- (xxix) SEQ ID NO:
- 85 is inserted into hsa-mir-221 (SEQ ID NO:
- 149);
- (xxx) SEQ ID NO:
- 86 is inserted into hsa-mir-222 (SEQ ID NO:
- 150);
- (xxxi) SEQ ID NO:
- 88 is inserted into hsa-mir-30e (SEQ ID NO:
- 152);
- (xxxii) SEQ ID NO:
- 92 is inserted into hsa-mir-423 (SEQ ID NO:
- 156);
- (xxxiii) SEQ ID NO:
- 93 is inserted into hsa-mir-519c (SEQ ID NO:
- 157);
- (xxxiv) SEQ ID NO:
- 96 is inserted into hsa-mir-92b (SEQ ID NO:
- 160);
- (xxxv) SEQ ID NO:
- 97, SEQ ID NO:
- 103, SEQ ID NO:
- 104 or SEQ ID NO:
- 109 is inserted into hsa-mir-302a (SEQ ID NO:
- 161, SEQ ID NO:
- 167, SEQ ID NO:
- 168 or SEQ ID NO:
- 173 respectively);
- (xxxvi) SEQ ID NO:
- 98, SEQ ID NO:
- 107, SEQ ID NO:
- 108 or SEQ ID NO:
- 111 is inserted into hsa-mir-302c (SEQ ID NO:
- 162, SEQ ID NO:
- 171, SEQ ID NO:
- 172 or SEQ ID NO:
- 175 respectively);
- (xxxvii) SEQ ID NO:
- 100 is inserted into hsa-mir-191 (SEQ ID NO:
- 164);
- (xxxviii) SEQ ID NO:
- 101 or SEQ ID NO:
- 102 is inserted into hsa-mir-30 (SEQ ID NO:
- 165 or SEQ ID NO:
- 166 respectively);
- (xxxix) SEQ ID NO:
- 105, SEQ ID NO:
- 106 or SEQ ID NO:
- 110 is inserted into hsa-mir-302b (SEQ ID NO:
- 169, SEQ ID NO:
- 170 or SEQ ID NO:
- 174 respectively);
- (xxxx) SEQ ID NO:
- 112, SEQ ID NO:
- 113 or SEQ ID NO:
- 114 is inserted into hsa-mir-375 (SEQ ID NO:
- 176, SEQ ID NO:
- 177 or SEQ ID NO:
- 178 respectively);
- (xxxxi) SEQ ID NO:
- 115, SEQ ID NO:
- 119, SEQ ID NO:
- 122 or SEQ ID NO:
- 125 is inserted into hsa-let-7b (SEQ ID NO:
- 179, SEQ ID NO:
- 183, SEQ ID NO:
- 186 or SEQ ID NO:
- 189 respectively);
- (xxxxii) SEQ ID NO:
- 116, SEQ ID NO:
- 120, SEQ ID NO:
- 123 or SEQ ID NO:
- 126 is inserted into hsa-let-7g (SEQ ID NO:
- 180, SEQ ID NO:
- 184, SEQ ID NO:
- 187 or SEQ ID NO:
- 190 respectively);
- and/or
- (xxxxiii) SEQ ID NO:
- 117, SEQ ID NO:
- 118, SEQ ID NO:
- 121, SEQ ID NO:
- 124 or SEQ ID NO:
- 127 is inserted into hsa-mir-98 (SEQ ID NO:
- 181, SEQ ID NO:
- 182, SEQ ID NO:
- 185, SEQ ID NO:
- 188 or SEQ ID NO:
- 191 respectively).
The polynucleotide sequence encoding an inhibitory RNA according to any preceding claim, which comprises a genomic sequence flanking both sides of a human pre-microRNA suitably wherein the polynucleotide sequence comprises:
- (i) SEQ ID NO:
- 35 (which comprises SEQ ID NO:
- 2 or SEQ ID NO:
- 19);
- (ii) SEQ ID NO:
- 36 (which comprises SEQ ID NO:
- 11 or SEQ ID NO:
- 28);
- (iii) SEQ ID NO:
- 37 (which comprises SEQ ID NO:
- 3 or SEQ ID NO:
- 20);
- (iv) SEQ ID NO:
- 38 (which comprises SEQ ID NO:
- 12 or SEQ ID NO:
- 29);
- (v) SEQ ID NO:
- 39 (which comprises SEQ ID NO:
- 9 or SEQ ID NO:
- 26);
- (vi) SEQ ID NO:
- 40 (which comprises SEQ ID NO:
- 8 or SEQ ID NO:
- 25);
- (vii) SEQ ID NO:
- 41 (which comprises SEQ ID NO:
- 10 or SEQ ID NO:
- 27);
- (viii) SEQ ID NO:
- 42 (which comprises SEQ ID NO:
- 13 or SEQ ID NO:
- 30);
- (iv) SEQ ID NO:
- 43 (which comprises SEQ ID NO:
- 4 or SEQ ID NO:
- 21);
- (x) SEQ ID NO:
- 44 (which comprises SEQ ID NO:
- 16 or SEQ ID NO:
- 33);
- (xi) SEQ ID NO:
- 45 (which comprises SEQ ID NO:
- 5 or SEQ ID NO:
- 22;
- (xii) SEQ ID NO:
- 46 (which comprises SEQ ID NO:
- 6 or SEQ ID NO:
- 23;
- (xiii) SEQ ID NO:
- 47 (which comprises SEQ ID NO:
- 1 or SEQ ID NO:
- 18;
- (xvi) SEQ ID NO:
- 48 (which comprises SEQ ID NO:
- 7 or SEQ ID NO:
- 24;
- (xv) SEQ ID NO:
- 49 (which comprises SEQ ID NO:
- 15 or SEQ ID NO:
- 32;
- (xvi) SEQ ID NO:
- 50 (which comprises SEQ ID NO:
- 14 or SEQ ID NO:
- 31;
- and/or
- (xvii) SEQ ID NO:
- 51 (which comprises SEQ ID NO:
- 17 or SEQ ID NO:
- 34).
A synthetic microRNA comprising or consisting of a sequence selected from any one of SEQ ID Nos:
- 1-17, 64-127 or functional variants or fragments thereof.
A synthetic pre-microRNA comprising or consisting of a sequence selected from any one of SEQ ID Nos:
- 18-34, 128-191 or functional variants or fragments thereof.
A synthetic microRNA adapted to target a messenger RNA transcribed from the B2M gene, the synthetic microRNA comprising a sequence according to any one of SEQ ID Nos:
- 1-17, 64-127 or a functional variant or fragment thereof, optionally wherein the synthetic microRNA comprises a sequence according to any one of SEQ ID NOs:
- 18-34, 128-191 or a functional variant or fragment thereof.
A cell comprising or expressing a sequence encoding a synthetic inhibitory RNA, preferably a synthetic microRNA, wherein the synthetic inhibitory RNA is adapted to target B2M.
A method of treating an autoimmune disease or cancer in a subject, preferably wherein the autoimmune disease is Type I diabetes, wherein the method comprises administering the cell or composition in a therapeutic amount to the subject.
A method of modifying the translation of a messenger RNA transcript derived from a protein coding gene and subsequent expression of a protein in a cell comprising introducing into the cell a DNA editing agent conferring a silencing specificity of a microRNA towards a target RNA of interest, wherein the microRNA is modified such that the sequence of the microRNA comprises any one of SEQ ID NOs:
- 1-17, 64-127 thereby modifying the translation of the messenger RNA transcript into a protein molecule.
Dependencies are inferred from the claim text.