siRNA targeting DNA-damage-inducible transcript 4 (DDIT4)

• US 7,655,788 B2
• Filed: 07/24/2007
• Issued: 02/02/2010
• Est. Priority Date: 11/14/2002
• Status: Active Grant

• Current Assignee: Thermo Fisher Scientific Incorporated
• Original Assignee: Dharmacon Incorporated (Revvity, Inc.)
• Inventors: Marshall, William, Khvorova, Anastasia, Reynolds, Angela, Read, Steven, Scaringe, Stephen, Leake, Devin
• Primary Examiner(s): McGarry; Sean
• Assistant Examiner(s): Gibbs; Terra Cotta
• Application Number:
  US11/880,775
• Publication Number:
  US 20080188647A1
• Time in Patent Office: 924 Days
• Field of Search: 536/23.1, 536/24.3, 536/24.33, 536/24.5, 514/44
• US Class Current: 536/24.5
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C12N 15/1048 SELEX
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 15/1136 against growth factors, growth regulators, cytokines, lymphokines or hormones
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2320/10 in screening processes
• CPC Class Codes: C12N 2320/11 for the determination of target sites, i.e. of active nucleic acids
• CPC Class Codes: C12Y 113/12007 Photinus-luciferin 4-monooxygenase (ATP-hydrolysing) (1.13.12.7), i.e. firefly-luciferase
• CPC Class Codes: C12Y 502/01008 Peptidylprolyl isomerase (5.2.1.8), i.e. cyclophilin
• CPC Class Codes: G16B 20/00 : ICT specially adapted for functional genomics or proteomics, e.g. genotype-phenotype associations
• CPC Class Codes: G16B 20/20 : Allele or variant detection, e.g. single nucleotide polymorphism [SNP] detection
• CPC Class Codes: G16B 20/50 : Mutagenesis

Abstract

Efficient sequence specific gene silencing is possible through the use of siRNA technology. By selecting particular siRNAs by rational design, one can maximize the generation of an effective gene silencing reagent, as well as methods for silencing genes. Methods, compositions, and kits generated through rational design of siRNAs are disclosed including those directed to DDIT4.

Claims

1. An siRNA molecule effective at silencing protein expression, wherein said siRNA molecule comprises a sense region and an antisense region, wherein said sense region and said antisense region together form a duplex region comprising 19-30 base pairs and said antisense region comprises a sequence that is the complement of SEQ. ID NO. 461.

2. The siRNA molecule of claim 1, wherein said antisense region and said sense region are each 19-25 nucleotides in length.

3. The siRNA molecule of claim 2, wherein said antisense region and said sense region are each 19 nucleotides in length.

4. The siRNA molecule of claim 1, wherein said antisense region is 100% complementary to a region of a target mRNA.

5. The siRNA molecule of claim 1, wherein said siRNA molecule comprises at least one overhang region, wherein said overhang region comprises six or fewer nucleotides.

6. The siRNA molecule of claim 1, wherein said siRNA molecule comprises no overhang regions.

7. The siRNA molecule of claim 1, wherein said siRNA molecule comprises at least one overhang region, wherein said overhang region comprises six or fewer nucleotides.

8. The siRNA molecule of claim 2, wherein said siRNA molecule comprises no overhang regions.

9. The siRNA molecule of claim 3, wherein said siRNA molecule comprises at least one overhang region, wherein said overhang region comprises six or fewer nucleotides.

10. The siRNA molecule of claim 3, wherein said siRNA molecule comprises no overhang regions.

11. An siRNA composition of between 19 and 30 base pairs in length that has 0-4 overhang regions, wherein said overhang regions are six or fewer bases in length, and wherein said composition comprises a sequence that is the complement of SEQ. ID. NO. 461.

12. The siRNA composition of claim 11 further comprising SEQ. ID. NO. 461.

13. The siRNA composition of claim 11, wherein said duplex is between 19 and 25 base pairs in length.

14. The siRNA composition of claim 13, wherein said duplex is 19 base pairs in length.

15. The siRNA composition of claim 11 further comprising at least one overhang region of six or fewer bases.

16. The siRNA composition of claim 12 further comprising at least one overhang region of six or fewer bases.

17. The siRNA composition of claim 11, wherein said siRNA comprises no overhang regions.

18. The siRNA composition of claim 12, wherein said siRNA comprises no overhang regions.

19. A chemically synthesized double stranded siRNA molecule that directs cleavage of a target mRNA via RNA interference, wherein:

a. each strand of said double stranded siRNA molecule is between 19 and 30 nucleotides in length; and

b. one strand of said double stranded siRNA molecule comprises a sequence that is the complement of SEQ. ID. NO. 461.

20. The chemically synthesized double stranded siRNA molecule of claim 19, wherein each strand of said double stranded siRNA molecule is 19 nucleotides in length.