Compositions and methods for modulation of LMNA expression
Disclosed herein are compounds, compositions and methods for modulating the expression of LMNA in a cell, tissue or animal. Also provided are methods of target validation. Also provided are uses of disclosed compounds and compositions in the manufacture of a medicament for treatment of diseases and disorders. Further provided are methods of identifying cis splicing regulatory elements of a selected mRNA using the disclosed compounds.
1. A single-stranded antisense oligonucleotide 12 to 30 nucleobases in length and at least 95% complementary to a nucleic acid molecule encoding human LMNA (SEQ ID NO: 4), wherein said antisense oligonucleotide comprises at least one 2′-O-(2-methoxyethyl) nucleotide, wherein said antisense oligonucleotide is a chimeric oligonucleotide comprising a first region comprising one or more deoxynucleotides and second and third regions flanking said first region, each comprising at least one 2′-O-(2-methoxyethyl) nucleotide, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 55 or 106.
2. The antisense oligonucleotide of claim 1 which is 16 to 20 nucleobases in length.
3. The antisense oligonucleotide of claim 1 , wherein said first region comprises 10 nucleobases and said second and third regions each comprise 5 nucleobases.
4. The antisense oligonucleotide of claim 1 further comprising a modified internucleoside linkage at each position.
5. The antisense oligonucleotide of claim 4 , wherein the modified internucleoside linkage at each position is a phosphorothioate.
6. The antisense oligonucleotide of claim 1 , wherein each cytosine is replaced with 5-methylcytosine.
7. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases of SEQ ID No: 106.
8. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide has a nucleobase sequence consisting of the sequence recited in any of the nucleobase sequences of SEQ ID NOs: 55 or 106.
9. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide has a nucleobase sequence consisting of the sequence recited in the nucleobase sequence of SEQ ID NO: 106.
10. The antisense oligonucleotide of claim 8 , further comprising a modified internucleoside linkage at each position.
11. The antisense oligonucleotide of claim 10 , wherein the modified internucleoside linkage at each position is a phosphorothioate.
12. The antisense oligonucleotide of claim 11 , wherein each cytosine is replaced with 5-methylcytosine.
13. A pharmaceutical composition comprising the antisense oligonucleotide of claim 1 and a pharmaceutically acceptable penetration enhancer, carrier or diluent.
14. A method of inhibiting expression of LMNA in cells or tissues, comprising contacting said cells or tissues with the antisense oligonucleotide of claim 1 .