Neurogenin3 and production of pancreatic islet cells
AssigneeThe Regents of the University of California
InventorMichael S. German
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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.
Claims29
A recombinant polypeptide comprising:
- a basic helix-loop-helix (bHLH) domain comprising an amino acid sequence that is at least 90% identical to a contiguous amino acid sequence of residues 82 to 138 of SEQ ID NO:2;
- and
- an overall amino acid sequence identity of at least 85% to a contiguous amino acid sequence of SEQ ID NO:2;
- wherein the recombinant polypeptide induces insulin gene transcription when expressed in a mammalian cell.
A recombinant polypeptide comprising an activation domain comprising an amino acid sequence that is at least 80% identical to amino acid residues 190-214 of SEQ ID NO:2, wherein the polypeptide induces gene transcription when expressed in a mammalian cell.
A recombinant polypeptide comprising:
- a basic helix-loop-helix (bHLH) domain comprising an amino acid sequence that is at least 90% identical to a contiguous amino acid sequence of residues 82 to 138 of SEQ ID NO:2;
- and
- an activation domain comprising an amino acid sequence that is at least 80% identical to amino acid residues 190-214 of SEQ ID NO:2, wherein the polypeptide induces gene transcription when expressed in a mammalian cell.
An isolated polypeptide comprising at least 85% amino acid sequence identity to the contiguous amino acid sequence of SEQ ID NO:2.
An isolated polypeptide comprising an amino acid sequence that is at least 85% identical to a contiguous amino acid sequence of residues 76-215 of SEQ ID NO:2.
An isolated polynucleotide encoding a neurogenin3 promoter, wherein the polynucleotide comprises a nucleotide sequence having an overall sequence identity of at least 80% to:
- a) a contiguous nucleotide sequence of nucleotides positioned at −105 to −158 of SEQ ID NO:1 relative to the transcription start site at nucleotide residue 2643 of SEQ ID NO:1;
- b) a contiguous nucleotide sequence of nucleotides positioned at-3728 to −3653 of SEQ ID NO:1 relative to the transcription start site at nucleotide residue 2643 of SEQ ID NO:1;
- c) a contiguous nucleotide sequence of nucleotides 2435 to 2643 of SEQ ID NO:1;
- d) a contiguous nucleotide sequence extending about 2.6 kb 5′ of nucleotide residue 2643 of SEQ ID NO:1;
- e) a contiguous nucleotide sequence extending about 5.7 kb 5′ of nucleotide residue 2643 of SEQ ID NO:1;
- or
- f) a contiguous nucleotide sequence of the sequence extending about 700 bp 5′ of nucleotide residue 719 of SEQ ID NO:3.
A method for producing a mammalian islet cell, the method comprising the steps of:
- introducing into a mammalian pancreatic cell a nucleic acid molecule encoding an islet transcription factor, which islet transcription factor is a neuroendocrine bHLH transcription factor, said introducing providing for expression of the islet transcription factor in the cell and for production of the islet cell phenotype in the cell.
A method for producing islet cells in a mammalian subject, the method comprising:
- administering a formulation to a pancreatic cell of a mammalian subject, the formulation comprising a nucleic acid molecule encoding and adapted for expression of an islet transcription factor, which islet transcription factor is a neuroendocrine bHLH transcription factor, said administering being in an amount sufficient for production of the islet transcription factor and production of islet cells in the pancreas of the subject.
A method of delivering insulin to the bloodstream of a mammalian subject, the method comprising:
- administering a formulation to a pancreatic cell of a mammalian subject, the formulation comprising a nucleic acid molecule encoding an islet transcription factor, which islet transcription factor is a neuroendocrine bHLH transcription factor, said administering being in an amount sufficient for production of the islet transcription factor at a level sufficient to induce production of beta cells, wherein the islet cells produce insulin, which insulin is delivered to the bloodstream of the subject.
Dependencies are inferred from the claim text.