RABBIT GENOME EDITING WITH ZINC FINGER NUCLEASES

• US 20110023140A1
• Filed: 07/23/2010
• Published: 01/27/2011
• Est. Priority Date: 12/04/2008
• Status: Abandoned Application

• Current Assignee: Sigma-Aldrich Company (Sigma-Aldrich Corporation)
• Original Assignee: Sigma-Aldrich Company (Sigma-Aldrich Corporation)
• Inventors: Buntaine, Brian, Cui, Xiaoxia, Bedell, Joseph
• Application Number:
  US12/842,208
• Publication Number:
  US 20110023140A1
• Time in Patent Office: Days
• US Class Current: 800/3
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2227/107 Rabbit
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A01K 67/0276 Knock-out vertebrates
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2800/80 Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113

Abstract

The present invention provides a genetically modified rabbit or cell comprising at least one edited chromosomal sequence. In particular, the chromosomal sequence is edited using a zinc finger nuclease-mediated editing process. The disclosure also provides zinc finger nucleases that target specific chromosomal sequences in the rabbit genome.

Claims

1. A genetically modified rabbit comprising at least one edited chromosomal sequence encoding a rabbit or human disease-related protein.

2. The genetically modified rabbit of claim 1, wherein the edited chromosomal sequence is inactivated, modified, or comprises an integrated sequence.

3. The genetically modified rabbit of claim 1, wherein the edited chromosomal sequence is inactivated such that no functional or even partially-functional rabbit or human disease-related protein is produced.

4. The genetically modified rabbit of claim 3, wherein inactivated chromosomal sequence comprises no exogenously introduced sequence(s).

5. The genetically modified rabbit of claim 3, further comprising at least one chromosomally integrated sequence encoding a functional rabbit or human disease-related protein.

6. The genetically modified animal of claim 1, wherein the rabbit or human disease is chosen from cardiovascular diseases;

ocular disease;

hypertriglyceridemia;

altered fat metabolism;

altered lipoprotein profile;

liver defects;

abnormal lipid metabolism;

diabetes and obesity;

autoimmune diseases;

immunodeficiency and combinations thereof.

7. The genetically modified rabbit of claim 1, wherein the rabbit is heterozygous or homozygous for the at least one edited chromosomal sequence.

8. The genetically modified rabbit of claim 1, wherein the rabbit is an embryo, a juvenile, or an adult.

9. The genetically modified rabbit of claim 1, wherein the protein is a human disease-related protein.

10. A rabbit embryo comprising at least one RNA molecule encoding a zinc finger nuclease that recognizes a chromosomal sequence encoding a rabbit or human disease-related protein, and, optionally, at least one donor polynucleotide comprising a sequence encoding a rabbit or human disease-related protein.

11. The rabbit embryo of claim 10, wherein the rabbit or human disease-related protein is chosen from cardiovascular diseases;

ocular disease;

hypertriglyceridemia;

altered fat metabolism;

altered lipoprotein profile;

liver defects;

abnormal lipid metabolism;

diabetes and obesity;

autoimmune diseases;

immunodeficiency and combinations thereof.

12. The rabbit embryo of claim 10, wherein the protein is a human disease-related protein.

13. A genetically modified rabbit cell, the cell comprising at least one edited chromosomal sequence encoding a rabbit or human disease-related protein.

14. The genetically modified cell of claim 13, wherein the edited chromosomal sequence is inactivated, modified, or comprises an integrated sequence.

15. The genetically modified cell of claim 13, wherein the edited chromosomal sequence is inactivated such that no functional rabbit or human disease-related protein is produced.

16. The genetically modified cell of claim 15, wherein the inactivated chromosomal sequence comprises no exogenously introduced sequence(s).

17. The genetically modified cell of claim 16, further comprising at least one chromosomally integrated sequence encoding a functional rabbit or human disease-related protein.

18. The genetically modified cell of claim 13, wherein the rabbit or human disease-related protein is chosen from cardiovascular diseases;

ocular disease;

hypertriglyceridemia;

altered fat metabolism;

altered lipoprotein profile;

liver defects;

abnormal lipid metabolism;

diabetes and obesity;

autoimmune diseases;

immunodeficiency and combinations thereof.

19. The genetically modified cell of claim 13, wherein the cell is heterozygous or homozygous for the at least one edited chromosomal sequence.

20. The genetically modified cell of claim 13, wherein the protein is a human disease-related protein.

21. A method for assessing the effect of an agent in a rabbit, the method comprising contacting a genetically modified rabbit comprising at least one edited chromosomal sequence encoding a rabbit or human disease-related protein with the agent, and comparing results of a selected parameter to results obtained from contacting a wild-type rabbit with the same agent, wherein the selected parameter is chosen from:

a) rate of elimination of the agent or its metabolite(s);

b) circulatory levels of the agent or its metabolite(s);

c) bioavailability of the agent or its metabolite(s);

d) rate of metabolism of the agent or its metabolite(s);

e) rate of clearance of the agent or its metabolite(s);

f) toxicity of the agent or its metabolite(s); and

g) efficacy of the agent or its metabolite(s).

22. The method of claim 21, wherein the agent is a pharmaceutically active ingredient, a drug, a toxin, biological active agent, or a chemical.

23. The method of claim 21, wherein the at least one edited chromosomal sequence is inactivated such that a functional rabbit or human disease-related protein is not produced, and wherein the animal further comprises at least one chromosomally integrated sequence encoding a functional rabbit or human disease-related protein.

24. The method of claim 21, wherein the rabbit or human disease is chosen from cardiovascular diseases;

ocular disease;

hypertriglyceridemia;

altered fat metabolism;

altered lipoprotein profile;

liver defects;

abnormal lipid metabolism;

diabetes and obesity;

autoimmune diseases;

immunodeficiency and combinations thereof.

25. The method of claim 21, wherein the rabbit is one of the species chosen from New Zealand White rabbit, Dutch rabbit, Flemish Giant rabbit, European Rabbit (Oryctolagus cuniculus) and any other strains thereof;

African Savanna Hare (Lepus victoriae);

Alaskan Hare (Lepus othus);

Amami Rabbit (Pentalagus furnessi);

Antelope Jackrabbit (Lepus alleni);

Arctic Hare (Lepus arcticus);

Black Jackrabbit (Lepus insularis);

Black-tailed Jackrabbit (Lepus californicus);

Broom Hare (Lepus castroviejoi);

Brush Rabbit (Sylvilagus bachmani);

Bunyoro Rabbit (Poelagus marjorita);

Burmese Hare (Lepus pequensis);

Brown Hare (Lepus capensis);

Chinese Hare (Lepus sinensis);

Corsican Hare (Lepus corsicanus);

Desert Cottontail (Sylvilagus audubonii);

Dice'"'"'s Cottontail (Sylvilagus dicei);

Eastern Cottontail (Sylvilagus floridanus);

Ethiopean Hare (Lepus fagani);

Ethiopean Highland Hare (Lepus starcki);

European Hare (Lepus europaeus);

Granada Hare (Lepus granatensis);

Hainan Hare (Lepus hainanus);

Hispid Hare (Caprolagus hispidus);

Indian Hare (Lepus nigricollis);

Jameson'"'"'s Red Rock Hare (Pronolagus randensis);

Japanese Hare (Lepus bracyurus);

Korean Hare (Lepus coreanus);

Marsh Rabbit (Sylvilagus palustris);

Mexican Cottontail (Sylvilagus cunicularius);

Mountain Cottontail (Sylvilagus nuttallii);

Mountain Hare (Lepus timidus);

Natal Red Rock Hare (Pronolagus crassicaudatus);

New England Cottontail (Sylvilagus transitionalis);

Omilteme Cottontail (Sylvilagus insonus);

Pygmy Rabbit (Brachylagus idahoensis);

Riverine Rabbit (Bunolagus monticularis);

San Jose Brush Rabbit (Sylvilagus mansuetus);

Scrub Hare (Lepus saxatilis);

Smith'"'"'s Red Rock Hare (Pronolagus rupestris);

Sumatra Short Eared Rabbit (Nesolagus netscheri);

Snowshoe Hare (Lepus americanus);

Swamp Rabbit (Sylvilagus aquaticus);

Tapeti (Sylvilagus brasiliensis);

Tehuantepec Jackrabbit (Lepus flavigularis);

Tolai Hare (Lepus tolai);

Tres Marias Cottontail (Sylvilagus graysoni);

Volcano Rabbit (Romerolagus diazi);

White-sided Jackrabbit (Lepus callotis);

White-tailed Jackrabbit (Lepus townsendii);

Woolly Hare (Lepus oiostolus);

Yarkand Hare (Lepus yarkandensis);

Yunnan Hare (Lepus comus) and other existing species.