Neurogenin 3 promoter

a promoter and neuronal growth factor technology, applied in the field of neuronal growth factor 3, can solve the problems of delayed treatment, increased complications, and premature heart attacks and strokes, and accounts for 15% of u.s. health care costs

Inactive Publication Date: 2004-08-05
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

0111] It should be noted that the invention contemplates the hNgn3 polypeptides and variants thereof described herein, as well as nucleic acids encoding such variants. When provided with guidance as to the amino acid sequence of a polypeptide and particularly wher

Problems solved by technology

Diabetes is also a major cause of premature heart attacks and stroke and accounts for 15% of U.S. health care costs.
Unfortunately, about one-half of the people in the U.S. affected by Type 2 diabetes are unaware that they have the disease.
Clinical symptoms associated with Type 2 diabetes may not become obvious until late in the disease, and the early signs are often misdiagnosed, causing a delay in treatment and increased complications.
Unfortunately, islet cell regeneration does not appear to occur when the islet cells alone are damaged, such as in type 1 diabetes.
Second, as discussed above, insulin production is impaired in individuals with diabetes.
In type 1 diabetes the impairment is caused by the destruction of the .beta.-cells, while in type 2 diabetes, insulin production is intact, but inadequate.
To date this has been largely unsucc

Method used

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Examples

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example 1

Detection of Ngn3 Expression in Murine Pancreas

[0260] Members of the basic helix-loop-helix (bHLH) family of transcription factors regulate growth and differentiation of numerous cell types. Insulin gene expression is activated by a heterodimeric complex of two bHLH proteins: a ubiquitously expressed (class A) protein and a cell-type-specific (class B) partner, NeuroD1 / BETA2. NeuroD1 / BETA2 is also important for .beta.-cell development. The targeted disruption of the NeuroD1 / BETA2 gene in mice leads to a marked reduction of the .beta.-cell mass at birth due to increased apoptosis of islet cells late in fetal development. There is no apparent defect, however, in .beta.-cell formation or insulin gene expression, despite the postulated importance of this factor in .beta.-cell differentiation.

[0261] Assuming that this modest phenotype reflected the redundant expression of closely related class B bHLH proteins in the endocrine pancreas, the inventors searched for additional members of the...

example 2

Isolation and Sequencing of a Human Ngn3 Polypeptide-Encoding Polynucleotide

[0263] A probe derived from a cloned fragment of the murine Ngn3 gene (Sommer et al., supra) was used to screen a human genomic library. This screen resulted in the isolation of the genomic sequence provided as SEQ ID NO:1 in the sequence listing. Based on mapping of the murine start site using 5' RACE of mouse fetal pancreatic RNA, the transcriptional start site in the human Ngn3-encoding sequence is at nucleotide residue 2643. The coding sequence is between nucleotide residues 3022-3663, with a stop site at 3664-3666. No introns are within the 5' untranslated region (UTR) or the coding sequence of SEQ ID NO:1.

[0264] The promoter of Ngn3 is of interest, particularly given that is it exceptionally well-conserved between mouse, rat, and human. Given the role of Ngn3 in pancreatic and islet cell development, the Ngn3 promoter is likely key to determining the number of islet cells in the mature pancreas. The re...

example 3

Isolation and Sequencing of a Murine Ngn3 Polypeptide-Encoding Polynucleotide and Promoter

[0265] The full-length murine Ngn3 sequence and its 5' flanking sequences, which included the murine Ngn3 promoter, were obtained by sequencing a previously obtained mouse genomic DNA fragment (Sommer, et al., supra). The murine Ngn3 sequence is provided in the Sequence Listing as SEQ ID NO:3, with the encoded polypeptide provided as SEQ ID NO:4. The transcriptional start site was determined using the 5' RACE method and confirmed using RNase protection with RNA from fetal mouse pancreas, and is at nucleotide residue 719; the coding sequence for murine Ngn3 begins at nucleotide residue 1093. The promoter comprises a region approximately 500 bp upstream of the transcription start site.

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Abstract

The present invention features polypeptides having activity of human neurogenin3 (hNgn3), and nucleic acid encoding such polypeptide. The invention also features use of islet transcription factors such as hNgn3 to facilitate production of pancreatic islet cells from progenitor cells, and to facilitate insulin delivery by production of islet cells so produced.

Description

[0001] This application is 1) a continuation-in-part of PCT application serial no. PCT / US02 / 11166, filed Mar. 20, 2002 designating the United States and published in English, and 2) a continuation-in-part of U.S. application Ser. No. 09 / 817, 360, filed Mar. 20, 2001, which application is a continuation-in-part of U.S. application Ser. No. 09 / 535,145, filed Mar. 24, 2000, which application is entitled to the benefit of U.S. Provisional Application Serial No. 60 / 128,180, filed Apr. 6, 1999. The disclosures of each of these applications are incorporated herein by reference in their entireties.[0002] The invention relates generally to the field of delivery of insulin to a subject by production of islet cells, particularly insulin producing beta cells, and to islet transcription factors useful in such delivery (e.g., human neurogenin3).[0003] Diabetes mellitus is the third leading cause of death in the U.S. and the leading cause of blindness, renal failure, and amputation. Diabetes is al...

Claims

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Application Information

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IPC IPC(8): A61K35/12A61K48/00C07H21/02C07H21/04C07K14/47C12N5/071C12N15/12
CPCA01K2217/05C12N2840/203A61K48/005A61K48/0058C07H21/02C07H21/04C07K14/47C07K14/4705C12N5/0676C12N2501/60C12N2799/022C12N2830/008C12N2830/42C12N2830/85A61K35/12
Inventor GERMAN, MICHAEL S.
Owner RGT UNIV OF CALIFORNIA
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