Use and production of CHD8+/− transgenic animals with behavioral phenotypes characteristic of autism spectrum disorder

Patent metadata

Field	Value
Publication number	US-11459557-B2
Application number	US-201715467949-A
Country	US
Kind code	B2
Filing date	Mar 23, 2017
Priority date	Sep 24, 2014
Publication date	Oct 4, 2022
Grant date	Oct 4, 2022

Abstract

Methods and compositions for inducing a plurality of mutations in transgenic Cas9 eukaryotes to model a neuronal disease or disorder. The invention further comprehends testing putative treatments with such models, e.g., testing putative chemical compounds that may be pharmaceutically relevant for treatment or gene therapy that may be relevant for treatment, or combinations thereof. The invention allows for the study of genetic diseases and putative treatments to better understand and alleviate a genetic disease or a condition, e.g., autism, autism-spectrum disease or disorder, obsessive-compulsive disorder, or psychiatric disorders.

First claim

What is claimed is:

1. A method for modeling a neuronal/nervous system related disease or disorder, comprising:
delivering a vector encoding a plurality of guides to a genetically-modified eukaryotic cell or a genetically-modified non-human animal,
wherein the genetically-modified eukaryotic cell or genetically-modified non-human animal comprises a chromosomally integrated Cas9 gene that is constitutively expressed, or a chromosomally integrated Cas9 gene capable of being inducibly or conditionally expressed to produce a Cas9 protein,
wherein, when the Cas9 gene and the plurality of guides are expressed in the cell or animal, the plurality of guides are capable of guiding the Cas9 protein to a plurality of neuronal cell-specific target loci, thereby introducing multiple neuronal cell-specific mutations or substitutions at the neuronal cell-specific target loci.

2. The method of claim 1 , wherein the neuronal cell-specific mutations or substitutions are or have been correlated to the neuronal/nervous system related disease or disorder.

3. The method of claim 1 , wherein each of the guides targets a different genetic locus associated with a neuronal cell/tissue-specific multigenic disease or disorder.

4. The method of claim 3 , wherein the multigenic disorder is a psychiatric disease or disorder.

5. The method of claim 3 , wherein the multigenic disorder is autism, an autism-spectrum disease or disorder, or obsessive compulsive disorder.

6. The method of claim 3 , wherein the multigenic disorder is a neuronal-associated developmental disease, epilepsy, anencephaly, or ancephalocele.

7. The method of claim 1 , wherein the guides target one or more neuronal/nervous system genes so as to introduce a loss-of-function mutation or a gain-of-function mutation.

8. The method of claim 1 , wherein the guides target one or more loci which are transcriptional control elements.

9. The method of claim 1 , wherein each of the guides targets a different genetic locus associated with a neuronal developmental pathway or lineage.

10. The method of claim 1 , wherein from 3 to 50 neuronal cell-specific mutations or substitutions in from 3 to 50 target loci are introduced.

11. The method of claim 1 , wherein the genetically-modified non-human animal is a non-human primate, a rodent, a canine, a bovine, an ovine, a mouse, a dog, a goat, a pig, a rabbit, a fowl or poultry.

12. The method of claim 1 , wherein the genetically modified non-human animal comprises neuronal cells that comprise a Cas9 gene that is constitutively, or capable of being inducibly, or conditionally expressed.

13. The method of claim 1 , wherein the plurality of guides comprises guides that target one or more of SHANK3, SAPAP3, MECP2, CHD8, DYRK1A, GRIN2B, KATNAL2, RIMS1, SCN2A, POGZ, ADNP, ARID1B, TBR1, NLRC5, STAC2, CRYBG3, SLC4A8, RAD54L2, MUC5B, MK167, NEB, MYH9, TANC2, PIK3R3, ACO1, CAPN9, CTNNA2, GRIN2A, ASH1L, LRP1, STAG1, and SYNGAP1.

14. The method of claim 1 , further comprising screening for therapeutic molecules or agents that alleviate a symptom of the neuronal/nervous system related disease or disorder, wherein screening for the therapeutic molecules or agents comprises:
treating the cell or the transgenic non-human animal with one of the therapeutic molecules or agents;
determining a change in the symptom; and
selecting a drug candidate based on the change in the symptom.

15. The method of claim 14 , wherein the cell or the genetically modified non-human animal exhibits a loss-of-function or gain-of-function for at least one genetic locus associated with the neuronal/nervous system related disease or disorder.

16. The method of claim 15 , wherein the neuronal/nervous system related disease or disorder is autism spectrum disorder (ASD) and the at least one genetic locus is CHD8.

17. The method of claim 14 , comprising treating the genetically modified non-human animal with one of the therapeutic molecules or agents, wherein the transgenic non-human animal is a loss-of-function or gain-of-function heterozygote or homozygote.

18. The method of claim 1 , further comprising delivering donor templates and a plurality of neuronal cell-specific mutations or substitutions.

19. The method of claim 1 , wherein the Cas9 gene is present in the cell or animal prior to the vector is delivered.

Assignees

• Broad Inst Inc
• Massachusetts Inst Technology

Inventors

• Platt Randall Jeffrey
• Zhang Feng
• Slaymaker Ian

Classifications

• A01K2217/072

  maintaining or altering function, i.e. knock in · CPC title

• A01K2227/105

  Murine · CPC title

• C12N2320/11

  for the determination of target sites, i.e. of active nucleic acids · CPC title

• C12N2320/32

  Special delivery means, e.g. tissue-specific · CPC title

• A01K2267/0306

  Animal model for genetic diseases · CPC title