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WO1999005266B1 - Trans-species nuclear transfer - Google Patents

Trans-species nuclear transfer

Info

Publication number
WO1999005266B1
WO1999005266B1 PCT/US1998/015387 US9815387W WO9905266B1 WO 1999005266 B1 WO1999005266 B1 WO 1999005266B1 US 9815387 W US9815387 W US 9815387W WO 9905266 B1 WO9905266 B1 WO 9905266B1
Authority
WO
WIPO (PCT)
Prior art keywords
cells
nuclear transfer
donor
hours
enucleated
Prior art date
Application number
PCT/US1998/015387
Other languages
French (fr)
Other versions
WO1999005266A3 (en
WO1999005266A2 (en
Inventor
Neal L First
Tanja Dominko
Maissam Mitalipoua
Original Assignee
Wisconsin Alumni Res Found
Neal L First
Tanja Dominko
Maissam Mitalipoua
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Wisconsin Alumni Res Found, Neal L First, Tanja Dominko, Maissam Mitalipoua filed Critical Wisconsin Alumni Res Found
Priority to AU85875/98A priority Critical patent/AU8587598A/en
Publication of WO1999005266A2 publication Critical patent/WO1999005266A2/en
Publication of WO1999005266A3 publication Critical patent/WO1999005266A3/en
Publication of WO1999005266B1 publication Critical patent/WO1999005266B1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/873—Techniques for producing new embryos, e.g. nuclear transfer, manipulation of totipotent cells or production of chimeric embryos
    • C12N15/877—Techniques for producing new mammalian cloned embryos
    • C12N15/8771—Bovine embryos

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Developmental Biology & Embryology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

A method of producing cloned nuclear transfer embryos from differentiated donor cells is described. The method includes culturing differentiated donor cells in low serum medium and using bovine oocytes as recipient oocytes for donor cells from different species. This method may be used to produce genetically identical animals and transgenic animals.

Claims

AMENDED CLAIMS[received by the International Bureau on 23 April 1999 (23.04.99); original claims 1-29 replaced by amended claims 1-15 (3 pages)]
1. A method of producing nuclear transfer embryos from donor cells of one species and recipient oocytes from another species comprising: inducing the donor cells to undergo G0 arrest; fusing said donor cell to an enucleated recipient oocyte of another species to create a nuclear transfer embryo; and activating said nuclear transfer embryo.
2. The method of claim 1 wherein said G0 arrest of donor cells is induced by culture in low serum medium.
3. The method of claim 1 wherein said donor cells are selected from the group consisting of embryonic derived cells, germ cells, somatic cells, and genetically modified cells.
4. The method of claim 1 wherein said enucleated recipient oocyte is an enucleated bovine recipient oocyte.
5. The method of claim 4 wherein said enucleated bovine recipient oocyte is selected from the group of bovine oocytes undergoing nuclear maturation within 16 hours of beginning in vitro culture.
6. The method of claim 1 wherein said enucleated bovine recipient oocyte and said donor cell are fused by electric pulse to form a nuclear transfer embryo.
7. The method of claim 1 wherein said fusion is performed 16-32 hours after the beginning of in vitro culture .
8. The method of claim 1 wherein said nuclear transfer embryo is activated by elevating intracellular calcium and the incubating with a serine threonine kinase inhibitor.
9. The method of claim 8 wherein intracellular calcium is elevated by incubation with ionomycin and the serine threonine kinase inhibitor is DMAP.
10. The method of claim 1 wherein said activation is 16-32 hours after the beginning of in vitro culture.
11. The method of claim 6 wherein said fusion is 16-52 hours after the beginning of in vitro culture.
12. An embryo produced by the method of claim 1.
13. A method of producing nuclear transfer embryos from a donor cell of one species and a bovine recipient oocyte comprising: culturing non-bovine donor cells selected from the group consisting of embryonic derived cells, somatic cells, germ cells, and genetically modified cells in low serum medium so that said donor cells are induced to arrest in the G0 stage of the cell cycle; selecting a bovine recipient oocyte which has completed nuclear maturation before 16 hours from the beginning of in vitro culture; enucleating said bovine recipient oocyte after 16-32 hours of in vitro culture; placing said donor cell under the zone pellucida of said enucleated oocyte so that said donor cell contacts said enucleated oocyte; fusing said donor cell with said enucleated oocyte by electric pulse at 16-32 hours after the beginning of in vitro culture to create a nuclear transfer embryo; and activating said nuclear transfer embryo by sequential incubation with ionomycin and 6-dimethylaminopurine at 16 to 32 hours after the beginning of in vitro culture.
14. The embryo produced by the process of claim 13.
15. A nuclear transfer embryo comprising cytoplasm and cell membrane from one species and differentiated cytoplasm, differentiated cell membrane, and nuclei derived from a differentiated cell of another species .
STATEMENT UNDER ARΗCLE 19
The original claims 1-29 have been amended, by canceling original claims 1-1 1 , 24, 25, and the remaining original claims 12-23, 26, 27 and 29 have been renumbered as claims 1-15. The claims after amendment are now directed solely to a method for producing nuclear transfer embryos from donor cells of one species and recipient oocytes from another species.
PCT/US1998/015387 1997-07-26 1998-07-24 Trans-species nuclear transfer WO1999005266A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU85875/98A AU8587598A (en) 1997-07-26 1998-07-24 Trans-species nuclear transfer

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US5310397P 1997-07-26 1997-07-26
US60/053,103 1997-07-26

Publications (3)

Publication Number Publication Date
WO1999005266A2 WO1999005266A2 (en) 1999-02-04
WO1999005266A3 WO1999005266A3 (en) 1999-04-15
WO1999005266B1 true WO1999005266B1 (en) 1999-09-23

Family

ID=21981947

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1998/015387 WO1999005266A2 (en) 1997-07-26 1998-07-24 Trans-species nuclear transfer

Country Status (2)

Country Link
AU (1) AU8587598A (en)
WO (1) WO1999005266A2 (en)

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US7696404B2 (en) 1996-08-19 2010-04-13 Advanced Cell Technology, Inc. Embryonic or stem-like cell lines produced by cross species nuclear transplantation and methods for enhancing embryonic development by genetic alteration of donor cells or by tissue culture conditions
EP0934403A4 (en) * 1996-08-19 2001-03-14 Univ Massachusetts Embryonic or stem-like cell lines produced by cross species nuclear transplantation
NZ506808A (en) * 1998-03-02 2003-12-19 Univ Massachusetts Embryonic or stem-like cell lines produced by cross- species nuclear transplantation
WO1999046982A1 (en) * 1998-03-16 1999-09-23 Relag Pty Ltd Porcine nuclear transfer
US7621606B2 (en) 2001-08-27 2009-11-24 Advanced Cell Technology, Inc. Trans-differentiation and re-differentiation of somatic cells and production of cells for cell therapies
JP2003503071A (en) * 1999-06-30 2003-01-28 アドバンスド セル テクノロジー、インコーポレイテッド Cytoplasmic transfer to dedifferentiated recipient cells
CN1304443A (en) * 1999-06-30 2001-07-18 黄禹锡 Method for producing clened cows
JP2003503045A (en) * 1999-06-30 2003-01-28 ワン，ウー−サク Clonal tiger production method using xenogeneic nuclear transfer technology
JP2003503044A (en) * 1999-06-30 2003-01-28 ワン，ウー−サク Method for producing human cloned embryos using interspecies nuclear transfer technology
NZ517609A (en) * 1999-09-14 2004-02-27 Univ Massachusetts Embryonic or stem-like cell lines produced by cross species nuclear transplantation and methods for enhancing embryonic development by genetic alteration of donor cells or by tissue culture conditions
CN1425064A (en) * 1999-12-20 2003-06-18 马萨诸塞大学 Embryonic or stem-like cells produced by cross species nuclear transplantation
CA2659945C (en) 2005-08-03 2014-12-16 Advanced Cell Technology, Inc. Improved methods of reprogramming animal somatic cells
WO2013010045A1 (en) 2011-07-12 2013-01-17 Biotime Inc. Novel methods and formulations for orthopedic cell therapy
DK2986729T3 (en) 2013-04-16 2018-10-29 Regeneron Pharma TARGETED MODIFICATION OF ROOT THROUGH
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WO2016081923A2 (en) 2014-11-21 2016-05-26 Regeneron Pharmaceuticals, Inc. METHODS AND COMPOSITIONS FOR TARGETED GENETIC MODIFICATION USING PAIRED GUIDE RNAs
US11326184B2 (en) 2014-12-19 2022-05-10 Regeneron Pharmaceuticals, Inc. Methods and compositions for targeted genetic modification through single-step multiple targeting
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RU2019125818A (en) 2017-02-27 2021-03-29 Регенерон Фармасьютикалс, Инк. MODELS OF RETINOSCHISIS ON ANIMALS OTHER THAN HUMAN
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US20190032156A1 (en) 2017-07-31 2019-01-31 Regeneron Pharmaceuticals, Inc. Methods and compositions for assessing crispr/cas-induced recombination with an exogenous donor nucleic acid in vivo
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Also Published As

Publication number Publication date
WO1999005266A3 (en) 1999-04-15
AU8587598A (en) 1999-02-16
WO1999005266A2 (en) 1999-02-04

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