DNA regulatory element for the expression of transgenes in neurons of a subject and uses thereof

Abstract

The present invention provides for a recombinant nucleic acid molecule comprising a region of a calcium-calmodulin dependent kinase IIα promoter operatively linked to a gene of interest. The region of a calcium-calmodulin dependent kinase IIα promoter may comprise an 8.5 kilobase nucleic acid sequence which corresponds to the nucleic acid sequence of ATCC Accession No.: ______, designated pMM281. The present invention also provides a human cell line which has been stably transformed by a recombinant nucleic acid molecule comprising a gene of interest operatively linked to a nucleic acid encoding a calcium-calmodulin dependent kinase IIα promoter region which has a nucleotide sequence corresponding to the sequence of ATCC Accession No. ______, designated pMM281. The present invention also provides for a transgenic nonhuman mammal whose germ or somatic cells contain a nucleic acid molecule which encodes a gene of interest under the control of a CaMKIIα promoter (ATCC Accession No. ______), introduced into the mammal, or an ancestor thereof, at an embryonic stage. Another embodiment of the present invention is a method of evaluating whether a compound is effective in treating symptoms of a neurological disorder in a subject which comprises: (a) administering the compound to the transgenic nonhuman mammal whose germ or somatic cells contain a nucleic acid molecule which encodes a gene of interest under the control of a CaMKIIα promoter, and (b) comparing the neurological function the mammal in step (a) with neurological function of the transgenic mammal in the absence of the compound, thereby determining whether the compound is effective in treating symptoms of the neurological disorder in a subject.

1 - 5 . (canceled) 
     
     
         6 . A human cell line which has been stably transformed by a recombinant nucleic acid molecule comprising a gene of interest operatively linked to a nucleic acid encoding a calcium-calmodulin dependent kinase IIα promoter region which has a nucleotide sequence corresponding to the sequence of ATCC Accession No. 98582. 
     
     
         7 . The human cell line of  claim 6 , wherein the gene of interest comprises an acalcinurin gene, a gene involved in brain function, a growth factor gene, an ion channel gene, a kinase gene, a neurotransmitter gene, a neurotrophic factor gene, a phosphatase gene, a recombinase gene, a reporter gene, a receptor gene, a transactivator transcription factor gene, a transcription factor gene. 
     
     
         8 . The cell line of  claim 6 , wherein the cell line is a human neuronal cell line. 
     
     
         9 . A transgenic nonhuman mammal whose germ or somatic cells contain a nucleic acid molecule which encodes a gene of interest under the control of a CaMKIIα promoter (ATCC-Accession No. 98582), introduced into the mammal, or an ancestor thereof, at an embryonic stage. 
     
     
         10 . The transgenic nonhuman mammal of  claim 9 , wherein the gene of interest comprises an acalcinurin gene, a gene involved in brain function, a growth factor gene, an ion channel gene, a kinase gene, a neurotransmitter gene, a neurotrophic factor gene, a phosphatase gene, a recombinase gene, a reporter gene, a receptor gene, a transactivator transcription factor gene, a transcription factor gene. 
     
     
         11 . The transgenic nonhuman mammal of  claim 9 , wherein the nucleic acid Molecule contains an appropriate piece of genomic clone DNA from the mammal designed for homologous recombination. 
     
     
         12 . A method of treating a neurological disorder in a subject which comprises administering to the subject an effective amount of the recombinant nucleic acid of claim  1  so as to express the gene of interest in the subject and thereby treat the neurological disorder. 
     
     
         13 . The method of  claim 13 , wherein the neurological disorder is amnesia, Alzheimer's disease, amyotrophic lateral sclerosis, a brain injury, cerebral senility, chronic peripheral neuropathy, a cognitive disability, a degenerative disorder associated with learning, Down's Syndrome, dyslexia, electric shock induced amnesia or amnesia. Guillain-Barre syndrome, head trauma, Huntington's disease, a learning disability, a memory deficiency, memory loss, a mental illness, mental retardation, memory or cognitive dysfunction, multi-infarct dementia and senile dementia, myasthenia gravis, a neuromuscular disorder, Parkinson's disease, Pick's disease, a reduction in spatial memory retention, senility, or Turret's syndrome. 
     
     
         14 . A method of evaluating whether a compound is effective in treating symptoms of a neurological disorder in a subject which comprises: (a) administering the compound to the transgenic nonhuman mammal of  claim 9 , and (b) comparing the neurological function the mammal in step (a) with neurological function of the transgenic mammal in the absence of the compound, thereby determining whether the compound is effective in treating symptoms of the neurological disorder in a subject. 
     
     
         15 . The method of  claim 14 , wherein the neurological disorder is amnesia, Alzheimer's disease, amyotrophic lateral sclerosis, a brain injury, cerebral senility, chronic peripheral neuropathy, a cognitive disability, a degenerative disorder associated with learning, Down's Syndrome, dyslexia, electric shock induced amnesia or amnesia, Guillain-Barre syndrome, head trauma, Huntington's disease, a learning disability, a memory deficiency, memory loss, a mental illness, mental retardation, memory or cognitive dysfunction, multi-infarct dementia and senile dementia, myasthenia gravis, a neuromuscular disorder, Parkinson's disease, Pick's disease, a reduction in spatial memory retention, senility, or Turret's syndrome. 
     
     
         16 . The method of  claim 14 , wherein the compound is an organic compound, a nucleic acid, a small molecule, an inorganic compound, a lipid, or a synthetic compound. 
     
     
         17 . The method of  claim 14 , wherein the mammal is a mouse, a sheep, a bovine, a canine, a porcine, or a primate. 
     
     
         18 . The method of  claim 14 , wherein the subject is a human. 
     
     
         19 . The method of  claim 14 , wherein the administration comprises intralesional, intraperitoneal, intramuscular or intravenous injection; infusion;
 liposome-mediated delivery; gene bombardment; topical, nasal, oral, anal, ocular or otic delivery.   
     
     
         20 . A method of evaluating whether a-compound is effective in treating symptoms of a neurological disorder in a subject which comprises: (a) contacting a human neuronal cell of the human neuronal cell line of  claim 6  with the compound, and (b) comparing the neuronal cell function of the neuronal cell in step (a) with neuronal cell function in the absence of the compound, thereby determining whether the compound is effective in treating symptoms of the neurological disorder.