Purified and isolated PIWI family genes and gene products and therapeutic and screening methods using same

• US 6,723,534 B2
• Filed: 06/04/2001
• Issued: 04/20/2004
• Est. Priority Date: 12/04/1998
• Status: Expired due to Fees

• Current Assignee: Duke University
• Original Assignee: Duke University
• Inventors: Lin, Haifan
• Primary Examiner(s): Nguyen, Dave Trong
• Application Number:
  US09/873,737
• Publication Number:
  US 20020076797A1
• Time in Patent Office: 1,051 Days
• Field of Search: 536/23.1, 536/23.5, 435/69.1, 435/320.1, 435/325
• US Class Current: 435/69.1
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07K 14/47 from mammals

Abstract

A family of isolated and purified proteins and nucleic acids are disclosed. Particularly, piwi family proteins and cDNAs encoding the same are disclosed. Recombinant host cells, recombinant nucleic acids, recombinant proteins and transgenic animals are also disclosed, along with methods of producing each. Isolated and purified antibodies to piwi family homologs, and methods of producing the same, are also disclosed. piwi family gene products are characterized as having activity in the growth, proliferation and self-renewing division of stem cells, and proliferation of primordial germ cells. Thus, therapeutic, screening, culturing and transgenic methods involving these activities are also disclosed.

Claims

1. An isolated and purified polynucleic acid encoding a biologically active piwi family polypeptide comprising an amino acid sequence as set forth in any of SEQ ID NOs:

2, 4 and 6.

2. The polynucleic acid of claim 1, further comprising a piwi family-encoding polynucleic acid sequence as set forth in any of SEQ ID NOs:

3 and 5.

3. An isolated and purified polynucleic acid encoding a biologically active piwi family polypeptide, wherein the polynucleic acid is at least about 75% identical to a sequence as set forth in any of SEQ ID NOS:

1, 3, and 5, and wherein the polypeptide comprises the PIWI box.

4. The polynucleic acid of claim 3, wherein the encoded polypeptide comprises a vertebrate piwi family polypeptide.

5. The polynucleic acid of claim 3, wherein the encoded polypeptide comprises a mammalian piwi family polypeptide.

6. The polynucleic acid of claim 3, wherein the encoded polypeptide comprises a human piwi family polypeptide.

7. The polynucleic acid of claim 3, further defined as a DNA segment.

8. A recombinant vector comprising the polynucleic acid of claim 3.

9. The recombinant vector of claim 8, wherein the vector is a recombinant expression vector.

10. The recombinant vector of claim 8 further defined as cony rising at least a 1,000 nucleotide long contiguous stretch of a polynucleic and sequence as essentially set forth in any of SEQ ID NOs:

1, 3 and 5.

11. A recombinant host cell comprising the recombinant vector of claim 8.

12. The recombinant host cell of claim 11, wherein the host cell is a procaryotic cell.

13. The recombinant host cell of claim 11, wherein the host cell is a eukaryotic cell.

14. A method of preparing a piwi family polypeptide, comprising:

transforming a cell with the polynucleic acid of claim 3 to produce a piwi family under conditions suitable for the expression of said polypeptide.