Claims for Patent: 7,271,313

Summary for Patent: 7,271,313

Title:	Presenilin-deficient mouse model of age-dependent neurodegeneration and cognitive loss
Abstract:	The present invention is directed to recombinantly engineered mice that are deficient in the expression of both presenilin-1 and presenilin-2. The mice exhibit characteristics of age-dependent cognitive impairments and neurodegeneration similar to those seen in Alzheimer\'s disease patients. This presenilin-deficient mouse model can be used to screen compounds capable of slowing, preventing or reversing the progression of cognitive impairments and neurodegeneration. The invention is also directed to the development of treatments for Alzheimer\'s disease based on augmentation or restoration of presenilin function in the brain. On the basis of the findings described herein, the invention is further directed to the development of assays to detect functional presenilin deficiency in human individuals, preferably through analysis of presenilin substrates, which may provide biomarkers useful in the diagnosis of Alzheimer\'s disease.
Inventor(s):	Shen; Jie (Boston, MA)
Assignee:	The Brigham and Women\'s Hospital, Inc. (Boston, MA)
Application Number:	10/409,672
Patent Claims:	1. A presenilin-deficient mouse made by a method comprising: (a) making a first recombinantly engineered mouse whose genomic DNA comprises: (i) homozygous modified endogenous presenilin-1 alleles in which one or more exons of each presenilin-1 allele are flanked by lox P elements in such a manner that, upon exposure to Cre recombinase, said one or more exons are excised from said genomic DNA; (ii) a DNA sequence element coding for Cre recombinase under the control of a CaM kinase II promoter; (b) producing or obtaining a second recombinantly engineered mouse, whose genomic DNA comprises homozygous modified endogenous presenilin-2 alleles in which one or more exons have been interrupted by the introduction of an exogenous sequence that prevents each presenilin-2 allele from expressing a functional protein; (c) crossbreeding said first recombinantly engineered mouse with said second recombinantly engineered mouse to produce a progeny mouse whose genomic DNA comprises a presenilin-1 allele with exons flanked by loxP elements as described above in paragraph (a)(i); a DNA sequence element coding for Cre recombinase as described in paragraph (a)(ii); and a presenilin-2 allele in which one or more exons have been interrupted as described in paragraph (b); (d) further breeding said progeny mouse to produce a presenilin-deficient mouse whose genomic DNA comprises homozygous presenilin-1 alleles with exons flanked by loxP elements as described in paragraph (a)(i); a DNA sequence element coding for Cre recombinase as described in paragraph (a)(ii); and homozygous presenilin-2 alleles in which one or more exons have been interrupted as described in paragraph (b); and wherein, at some time after birth and prior to death, said presenilin-deficient mouse comprises within the genome of cells having CaM kinase II promoter activity a homozygous deletion within its presenilin-1 alleles such that, compared to a wild type mouse, said presenilin-deficient mouse is: (i) deficient in presenilin-1 in those cells where the CaM kinase II promoter is active; (ii) does not express presenilin-2; and (iii) exhibits age-dependent synaptic and neuronal degeneration and age-related memory impairment. 2. The presenilin-deficient mouse of claim 1, wherein said loxP elements are located in introns 1 and 3 of said presenilin-1 alleles. 3. The presenilin-deficient mouse of claim 2, wherein: (a) there is one loxP element in said intron 1; (b) said intron 3 further comprises a marker DNA sequence coding for a hygromycin resistance protein and coding for thymidine kinase, both of which are under the control of a CMV promoter; (c) there are two loxP elements in intron 3, one lying 5' to said marker DNA sequence and one lying 3' to said marker DNA sequence. 4. The presenilin-deficient mouse of claim 3, wherein said exogenous sequence that prevents said presenilin-2 alleles from producing a functional protein is in exon 5 of said presenilin-2 alleles. 5. The presenilin-deficient mouse of claim 3, wherein said exogenous sequence comprises a neomycin resistance gene under the control of an exogenous promoter. 6. The presenilin-deficient mouse of claim 1 wherein: a) said presenilin-deficient mouse is viable for at least six months after birth, and b) said presenilin-deficient mouse makes normal levels of presenilin-1 up until at least the time of its birth. 7. A transgenic mouse useful in the making of the presenilin-deficient mouse of claim 1, comprising within its genomic DNA: (a) a presenilin-1 gene in which one or more exons of said gene are flanked by loxP elements in such a manner that, upon exposure to Cre recombinase, said one or more exons are excised from said genomic DNA; and (b) a DNA sequence element coding for Cre recombinase under the control of a CaM kinase II promoter. 8. The transgenic mouse of claim 7, wherein said loxP elements are in intron 1 and intron 3 of said presenilin-1 gene. 9. The transgenic mouse of claim 7, wherein: (a) there is one loxP element in intron 1; and (b) there is at least one loxP element in intron 3.

Details for Patent 7,271,313

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2039-02-26
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2039-02-26
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2039-02-26