Methods and compositions for expressing polynucleotides specifically in smooth muscle cells in vivo
AssigneeSetagon, Inc.
InventorsGary K. Owens, Ichiro Manabe
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Abstract
The present invention generally relates to promoters, enhancers and other regulatory elements of smooth muscle cells (“SMC”). The invention more particularly relates to methods for the targeted knockout, or over-expression, of genes of interest within smooth muscle cells or within a subtype of smooth muscle cells. The invention further relates to methods of conferring polynucleotide expression in vivo specifically in smooth muscle cells or in subtypes of smooth muscle cells.
Claims28
An isolated polynucleotide comprising a smooth muscle cell myosin heavy chain (SM-MHC) promoter/enhancer, wherein the enhancer comprises nucleotides 5638-5860 of SEQ ID NO:16 or nucleotides 6862-7100 of SEQ ID NO:17, and the promoter comprises a heterologous TATA box or transcription initiation site, and wherein the promoter/enhancer initiates expression in a smooth muscle cell in vivo when introduced into an animal.
An isolated polynucleotide comprising a smooth muscle cell myosin heavy chain (SM-MHC) promoter/enhancer, wherein the promoter/enhancer sequence comprises SEQ ID NO:16 or SEQ ID NO:17, wherein:
- a CArG2 motif is mutated and the promoter is expressed in mesenteric artery, airway, stomach, intestine and bladder but not aorta, coronary artery or vena cava when introduced into an animal;
- or
- the intronic CArG motif at positions 5815-5825 of SEQ ID NO:16 or 7046-7056 of SEQ ID NO:17 is mutated and the promoter is expressed in coronary artery, mesenteric artery, vena cava, airway, stomach, intestine and bladder but not aorta when introduced into an animal.
An isolated polynucleotide comprising a smooth muscle cell myosin heavy chain (SM-MHC) promoter/enhancer, wherein the promoter/enhancer sequence comprises:
- nucleotides 1 to 9500 and 11,700 to 13,700 of SEQ ID NO:16 and does not comprise the intervening nucleotides 9501-11699 of SEQ ID NO:
- 16;
- or
- nucleotides 1 to 6700 and 9,500 to 15,800 of SEQ ID NO:16 and does not comprise the intervening nucleotides 6701-9499 of SEQ ID NO:16;
- and
- wherein the promoter/enhancer comprises a mutated or unmutated CArG2 or intronic CArG motif and the promoter/enhancer initiates expression in pulmonary vascular and airway smooth muscle cells in vivo when introduced into an animal.
Dependencies are inferred from the claim text.