1. A one-piece squeeze and turn child resistant closure for a container having a threaded neck and a top shoulder having a top surface with at least one locking recessed annulus slot or groove, the closure comprising a sidewall of a rigid material, the sidewall further comprising at least one flexible panel wherein, when depressed, allows the unscrewing of the closure from the bottle, wherein each flexible panel includes at least one vertical tab that engages a corresponding locking recessed slot or groove of the container.
2. The one-piece closure of claim 1 further comprising at least two flexible panels.
3. The one-piece closure of claim 2 comprising two flexible panels located at opposite sides of the bottle.
4. The one-piece closure of claim 1 further comprising at least one vertical stiffening rib supporting the inside diameter of the sidewall.
5. The one-piece closure of claim 1 further comprising at least one stiffening rib supporting the at least one vertical tab.
6. The one-piece closure of claim 1 further comprising a plug seal to plug an opening in the container.
7. A child resistant package comprising a container and a one-piece squeeze and turn child resistant closure for the container; the container having a threaded neck and a top shoulder having a top surface with at least one locking annulus recessed slot or groove, the closure comprising a sidewall of a rigid material, the sidewall further comprising at least one flexible panel wherein, when depressed, allows the unscrewing of the closure from the container, wherein each movable panel includes a vertical tab that engages a corresponding locking recessed slot or groove of the container.
8. The package of claim 7 wherein the at least one annulus slot or groove in the bottle prevent removal of the closure without first depressing the flexible panel.
9. The package of claim 7 wherein the closure further comprises at least two flexible panels.
10. The package of claim 9 wherein the closure further comprises two flexible panels located at opposite sides of the bottle.
11. The package of claim 7 wherein the closure further comprises at least one vertical stiffening rib supporting the inside diameter of the sidewall.
12. The package of claim 7 wherein the closure further comprising at least one stiffening rib supporting the at least one vertical tab.
13. The package of claim 7 wherein the closure further comprises a plug seal to plug an opening in the container.
14. The package of claim 7 wherein the container further comprises at least one finger grip.
15. The package of claim 7 wherein the outside diameter of the closure matches the outside diameter of the container.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A chimeric construct comprising at least (i) a smooth muscle cell (SMC)-specific promoter operably linked with at least (ii) a muscle-specific enhancer;
wherein the promoter and enhancer originate from different genes, different species, or different genes and species;
said promoter is selected from the group consisting of
a SMC-specific promoter comprising the 445 base pair region of the mouse SM22\u03b1 promoter set forth in SEQ ID NO: 1 from positions 1 to 445; and,
a SMC-specific promoter comprising the 445 base pair region of the human SM22\u03b1 promoter set forth in SEQ ID NO: 14 from positions 1 to 445;
and wherein said muscle-specific enhancer is selected from the group consisting of enhancers of the genes encoding: a rabbit myosin and a human creatine kinase.
2. The chimeric construct according to claim 1, wherein said muscle-specific enhancer is from the rabbit myosin heavy chain encoding gene.
3. The chimeric construct according to claim 2, wherein said muscle-specific enhancer comprises the sequence of nucleotides located 1332 to 1225 nucleotides upstream of the transcription initiation site of the rabbit myosin heavy chain encoding gene as set forth in SEQ ID NO: 2.
4. The chimeric construct according to claim 1, wherein said muscle-specific enhancer comprises the sequence of nucleotides located 919 to 711 nucleotides upstream of the transcription initiation site of the human creatine kinase gene as set forth in SEQ ID NO: 3.
5. An expression cassette comprising a gene of interest placed under the control of a chimeric construct according to claim 1, allowing its expression in a target cell.
6. The expression cassette according to claim 5, wherein said gene of interest encodes a polypeptide having growth suppressive activities on SMCs.
7. The expression cassette according to claim 6, wherein said polypeptide is selected from the group consisting of IFN\u03b2, IFN\u03b3, nitric oxide synthase eNOS, Fas ligand, heme oxygenase, interleukin-10 and heparin-binding VEGF.
8. The expression cassette according to claim 5, wherein said expression cassette further comprises one or more exons or portions of exons.
9. The expression cassette according to claim 8, which comprises the portion of the first non-coding exon of the mouse SM22\u03b1 gene extending from position 1 to position 65 downstream from its transcription initiation site as set forth in SEQ ID NO: 1 from position 446 to position 510 or the portion of the first non-coding exon of the human SM22\u03b1 gene extending from position 1 to position 63 downstream from its transcription initiation site as set forth in SEQ ID NO: 14 from position 446 to position 508.
10. A vector comprising the expression cassette according to claim 5.
11. The vector according to claim 10, wherein said vector is a viral vector derived from a virus selected from the group consisting of herpes viruses, cytomegaloviruses, foamy viruses, lentiviruses, Semliki forrest virus, AAV (adeno-associated virus), poxviruses, retroviruses and adenoviruses.
12. The vector according to claim 11, wherein said viral vector is a replication-defective adenoviral vector.
13. A method for the preparation of viral particles allowing the SMC-specific expression of a gene of interest in a target cell, said method comprising the steps of:
a) introducing the viral vector of claim 11 in a permissive cell line;
b) culturing the permissive cell line obtained in step a) for an appropriate period of time and under suitable conditions to allow the production of said viral particles;
c) recovering said viral particles from the cell culture; and
d) optionally, purifying the recovered viral particles.
14. A viral particle comprising the vector according to claim 10.
15. An isolated vascular smooth muscle cell comprising the expression cassette according to claim 5.
16. A method for specific expression of a gene of interest in SMCs in vitro comprising introducing the expression cassette according to claim 5 into said SMCs.