What is claimed is:
1. A method of inhibiting expression of a target allele in a cell comprising at least two different alleles of a gene, the method comprising administering to the cell an siRNA specific for the target allele.
2. The method of claim 1, wherein the target allele is correlated with a disorder associated with a dominant gain of function mutation.
3. The method of claim 2, wherein the disorder is selected from the group of amyotrophic lateral sclerosis, Huntington’s disease, Alzheimer’s disease, and Parkinson’s disease.
4. A method of treating a subject having a disorder correlated with the presence of a dominant gain of function mutant allele, the method comprising administering to the subject a therapeutically effective amount of an siRNA specific for the mutant allele.
5. The method of claim 4, wherein the siRNA is targeted to the gain of function mutation.
6. The method of claim 4, wherein the disorder is selected from the group of amyotrophic lateral sclerosis, Huntington’s disease, Alzheimer’s disease, and Parkinson’s disease.
7. The method of claim 4 wherein the disease is amyotrophic lateral sclerosis.
8. The method of claim 7 wherein the allele is SOD1.
9. The method of claim 8, wherein the mutant allele comprises a point mutation.
10. The method of claim 8, wherein the point mutation is a guanine: cytosine mutation.
11. The method of claim 8, wherein the mutation is G256C.
12. The method of claim 8, wherein the mutation is G281C.
13. The method of claim 8 wherein the siRNA comprises a sequence as set forth in FIG. 1A.
14. An siRNA comprising a sequence as set forth in FIG. 1A.
15. A p10 mutant siRNA comprising the sequence as set forth in FIG. 1A.
16. A p9 mutant siRNA comprising the sequence as set forth in FIG. 1A.
17. A G93A SOD1 shRNA comprising the sequence as set forth in FIG. 3A.
18. An expression construct comprising the shRNA of claim 13.
19. A therapeutic composition comprising the siRNA of claim 10-11, and a pharmaceutically acceptable carrier.
20. A therapeutic composition comprising the shRNA of claim 13, and a pharmaceutically acceptable carrier.
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 single use hypodermic syringe with a retracting needle system comprising:
a cylindrical body element for containing a fluid;
a needle assembly comprising a needle holding member and a hollow needle, the needle holding member repositionable between an injecting position wherein the needle projects axially outward from the body element and a pre-use and post-use position wherein the needle is retracted and contained completely within the body element;
a plunger rod reciprocally received into the body element forming a fluid chamber between the cylindrical body element, the plunger and the needle holding member,
and whereby positive and negative pressure forces are generated by adjustment of the plunger within the body element whereby fluid is drawn into, and expelled from, the body element through the hollow needle;
a retraction mechanism comprising:
a spring between the needle holding member and the axial end of the cylindrical body element through which the needle member projects and which generates an axial pressure on the needle holding member;
at least one retaining member engaging the needle holding member and retaining the needle holding member against the axial force of the spring;
at least one activating member within the needle holding member responding to positive fluid pressure within the fluid chamber to release the retaining member from the needle holding member;
whereby the needle is automatically deployed into the injecting position by depression of the plunger prior to drawing fluid into the fluid chamber and is automatically retracted within the body element at the completion of an injection stroke of the plunger and which retraction renders the syringe safe for handling and disposal.
2. The hypodermic syringe of claim 1 wherein said activating member comprises an activating ring with pistons attached cooperating with an activating release slide, whereby positive fluid pressure within the fluid chamber pushes the activating ring piston against the activating release slide which in turn releases the retaining clip from the needle holding member.
3. The hypodermic syringe of claim 1 wherein the spring is conical helical spring.
4. The hypodermic syringe of claim 3 wherein the spring is a twin conical helical spring.
5. The hypodermic syringe of claim 4 wherein the twin conical helical spring includes a spring separation member.
6. The hypodermic syringe of claim 1 wherein the spring are received between a pair of spring retaining cups.
7. The hypodermic syringe of claim 6 wherein one of spring retaining cups is integrally formed with the needle holding member.
8. The hypodermic syringe of claim 1 wherein a plurality of activation members are equidistantly spaced around the needle holding member, each activation member cooperating with a corresponding retaining member.
9. The hypodermic syringe of claim 8 wherein a slide member is located between each activation member and each corresponding retaining member.
10. The hypodermic syringe of claim 5 wherein the twin conical spring and the spring separation member cooperate to deflect the hollow needle from axial alignment when in the post-use position.
11. A single use hypodermic syringe with a retracting needle system comprising:
a cylindrical body element for containing a fluid;
a needle assembly comprising a needle holding member and a hollow needle, the needle holding member repositionable between an injecting position wherein the needle projects axially outward from the body element and a pre-use and post-use position wherein the needle is retracted and contained completely within the body element;
a plunger reciprocally received into the cylindrical body element forming a fluid chamber between the cylindrical body element and the needle holding member, and whereby positive and negative pressure forces are generated by adjustment of the plunger within the body element whereby fluid is drawn into, and expelled from, the cylindrical body element through the hollow needle;
a retraction mechanism comprising:
a spring between the needle holding member and the axial end of the cylindrical body element through which the needle member projects and which generates an axial force on the needle holding member, wherein the spring is a twin conical helical springs and include a spring separation member and is received between a pair of spring retaining cups, both spring retaining cups being integrally formed with the needle holding member;
at least one retaining member engaging the needle holding member and retaining the needle holding member against the axial force of the spring;
a plurality of activating members within the needle holding member equidistantly spaced around the needle holding member moulded on a ring, responding to positive fluid pressure within the fluid chamber to release a corresponding retaining member from the needle holding member, and a slide member located between each activation member and each corresponding retaining member;
whereby the needle is automatically deployed into the injecting position by depression of the plunger prior to drawing fluid into the fluid chamber and is automatically retracted within the cylindrical body element at the completion of an injection stroke of the plunger and which retraction renders the syringe safe for handling and disposal.
12. A needle retraction mechanism for a single use hypodermic syringe having a cylindrical body element for containing a fluid and a plunger reciprocally received into the body element forming a fluid chamber between the cylindrical body element and the needle holding member, and whereby positive and negative pressure forces are generated by adjustment of the plunger within the cylindrical body element whereby fluid is drawn into, and expelled from, the cylindrical body element through the hollow needle, comprising:
a needle assembly comprising a needle holding member and a hollow needle, the needle holding member repositionable between an injecting position wherein the needle projects axially outward from the body element and post-use position wherein the needle is retracted and contained completely within the cylindrical body element;
a spring between the needle holding member and the axial end of the cylindrical body element through which the needle member projects and which generates an axial force on the needle holding member;
at least one retaining member engaging the needle holding member and retaining the needle holding member against the axial force of the spring;
at least one activating member within the needle holding member responding to positive fluid pressure within the fluid chamber to release the retaining member from the needle holding member comprising:
an activating ring piston cooperating with an activating release slide, whereby positive fluid pressure within the fluid chamber pushes the activating ring piston against the activating release slide which in turn releases the retaining clip from the needle holding member.
13. The needle retraction mechanism of claim 12 wherein the spring is a conical helical spring.
14. The needle retraction mechanism of claim 13 wherein the spring is a conical helical spring.
15. The needle retraction mechanism of claim 14 wherein the twin conical helical spring includes a spring separation member.
16. The needle retraction mechanism of claim 12 wherein the spring is received between a pair of spring retaining cups.
17. The needle retraction mechanism of claim 16 wherein both spring retaining cups are integrally formed with the needle holding member.
18. The retraction mechanism of claim 12 wherein a plurality of activation members are equidistantly spaced around the needle holding member moulded on a ring, the activation member cooperating with a corresponding retaining member.
19. The needle retraction mechanism of claim 18 wherein a slide member is located between each activation member and each corresponding retaining member.
20. A needle retraction mechanism of claim 15 wherein the twin conical spring and the spring separation member cooperate to deflect the hollow needle from axial alignment when in the post-use position.