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WO2008017234A1 - Procédé de transfert nucléaire de cellules - Google Patents

Procédé de transfert nucléaire de cellules Download PDF

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Publication number
WO2008017234A1
WO2008017234A1 PCT/CN2007/002239 CN2007002239W WO2008017234A1 WO 2008017234 A1 WO2008017234 A1 WO 2008017234A1 CN 2007002239 W CN2007002239 W CN 2007002239W WO 2008017234 A1 WO2008017234 A1 WO 2008017234A1
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WO
WIPO (PCT)
Prior art keywords
oocyte
nuclear transfer
haplotype
cell
nuclear
Prior art date
Application number
PCT/CN2007/002239
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English (en)
Chinese (zh)
Inventor
Yitao Zeng
Fei Jiao
Fanyi Zeng
Shuzhen Huang
Original Assignee
Shanghai Jiao Tong University Affiliated Children's Hospital
Shanghai Tao Tao Engineering Co., Ltd
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 Shanghai Jiao Tong University Affiliated Children's Hospital, Shanghai Tao Tao Engineering Co., Ltd filed Critical Shanghai Jiao Tong University Affiliated Children's Hospital
Publication of WO2008017234A1 publication Critical patent/WO2008017234A1/fr

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    • 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

Definitions

  • the invention belongs to the field of bioengineering technology, and in particular to a method of nuclear transfer. Background technique
  • Somatic cell nuclear transfer (SCNT) technology has been widely used in the production of cloned and transgenic animals, and cloned animals of various species have been introduced (Chesne P., Adenot PG., Viglietta C., et al. Nat). Biotechnol. 2002, 20: 366-369; Woods GL" White L., Vanderwall DK” et al. Science 2003, 301: 1063 ).
  • cloning technology has unique advantages, this is Because the cloning technology advances the screening work to the cellular level, the breeding cycle of the elite animals is greatly shortened. Currently, low efficiency is a bottleneck that limits the technology.
  • Mitochondrion is not only the most abundant organelle in cytoplasm, but more importantly, due to the high mutation rate of mitochondria, it causes different mitochondrial DNA (mtDNA) sequences among different populations and different individuals in the same population. Haplotypes, this difference can cause differences in biological traits, such as differences in milk production and fertility in dairy cows (Tamassia M., Heyman Y., Lavergne Y., et al. Reproduction 2003, 126: 629) - 637; Sutrno, Cummins JM., Greeff J., et al. Theriogenology 2002, 57: 1603-1610; Mannen H., Kojima T" Oyama K., et al. J. Anim. Sci. 1998, 76: 36 -41 ) 0
  • haplotypes in the field of animal reproduction have increasingly become research hotspots.
  • the traditional concept of haplotype refers to the pattern of specific fragments produced by digestion of a specific fragment with a specific restriction endonuclease.
  • IVP in vitro embryonic production
  • somatic cell nuclear transfer indicate that in vitro embryo development is significantly affected by the maternal mtDNA haplotype and is associated with differences in oocyte ATP and mtDNA copy number (Tamassia M., Nuttinck F. Reynier ⁇ ,, et al. Biol. Reprod. 2004, 71: 697-704; Bruggerhoff K., Zakhartchenko V., Wenigerkind H., et al. Biol. Reprod. 2002, 66: 367-373; Hiendleder S. , Prelle K., Bruggerhoff K., et al. Biol. Reprod. 2004, 70: 1196-1205).
  • oocytes are beneficial to the development of cloned embryos, and may be related to the development of certain mtDNA haplotypes in the cytoplasm, which are more suitable for embryonic development.
  • the compatibility of mtDNA haplotypes may play an important role in facilitating the survival of cloned embryos of a particular mitochondrial type.
  • An object of the present invention is to provide a method for nuclear transfer which comprises nuclear transfer of a donor cell and a recipient oocyte of a non-human mammal having the same haplotype DNA haplotype.
  • the inventors performed haplotype analysis of mitochondrial DNA of experimental animals by PCR-RFLP technique, and classified mitochondrial DNA haplotypes of different experimental animals into four types, and then utilized the same haplotype nucleus.
  • the nuclear transfer of the cells and the recipient oocytes revealed that the nuclear transfer with the same haplotype can effectively improve the developmental ability of the reconstructed embryo.
  • the nuclear transfer method of the present invention comprises the following steps:
  • a selecting a non-human mammal of a mitochondrial DNA haplotype, and using it as a dermal fibroblast or a cumulus cell as a donor cell;
  • the nuclei of the nuclear donor cells are transferred into the cytoplasm of the enucleated oocyte, and the nuclei are integrated into the oocyte to form a reconstructed embryo.
  • the same type (the same mitochondrial DNA haplotype) cell nuclear transfer method can effectively improve the fusion rate, cleavage rate and blastocyst rate of the reconstructed embryo, compared with the heterotypic (mitochondrial DNA haplotype) nuclear transfer, blastocyst rate Increased by about 1.5 times.
  • PCR-RFLP polymerase chain reaction-restriction fragment length polymorphism
  • H2 S: 5'-TTATCCGTTGGTCTTAGGAA-3';
  • H3 S: 5,-TTATCACAATCCAGAACTGAC-3,;
  • H4 S: 5'-TGTGCATGTGACACGTATCC-3';
  • cycle conditions are: 94 °C 30s, 58 °C 30s, 72 °C lmin, 35 cycles; the last 72 °C extension for 10 min.
  • H2 94 °C pre-denaturation 5 min, enter the cycle; cycle conditions are: 94 °C lmin, 56 °C lmin, 72 °C 4min, 35 cycles; the last 72 °C extension for 10 min.
  • cycle conditions are: 94 °C lmin, 56. C lmin, 72 ° C for 4 min, 35 cycles; last 72 ° C for 10 min.
  • H4 94 °C pre-denaturation for 5 min, enter the cycle;
  • the cycle conditions are: 94 °C lmin, 56 °C lmin, 72 °C 5 min, 35 cycles; the last 72 °C extension for 10 min.
  • the obtained PCR amplification product was purified by PCR product purification kit (Takara) product specification (product number DV807A).
  • the purified amplification products were digested with restriction endonucleases, respectively, as follows:
  • Bamin was digested overnight at 30 °C, and the rest were digested overnight at 37 ⁇ .
  • the digested product was electrophoresed (0.8 ⁇ 2% agarose, 120V, lh) for further analysis.
  • Young bovine fibroblasts or cumulus cells classified by the above mitochondrial DNA haplotype were cultured in DMEM/F12 (Gibco, product number 11039-021, respectively) containing 10% FBS.
  • the mature 20h hour COCs (see Example 3) were placed in calcium-free magnesium-containing DPBS containing 0.5% hyaluronidase, digested for 1 ⁇ 2 min, and repeatedly sucked with a suction tube to remove the outer layer loose and expand. Cumulus cells.
  • the cells were suspended in DMEM/F12+ 10% FBS, gently pipetted, and the cells were thoroughly blown off, and inoculated into a 25 cm 2 flask (inoculation density of about IX 10 5 , containing DMEM/F12 + 10% FBS medium 5 ml), at 38.5 Incubate in an incubator at °C, 5% C0 2 , saturated humidity.
  • Cells cultured for 1 to 5 passages are used as donor cells for nuclear transfer.
  • the culture solution was replaced with a culture solution containing 0.5% FBS and starved for 2 to 3 days to induce the cells to enter the G0/G1 phase.
  • Lactic acid (Sigma L-7900) 14 ⁇ 1
  • BSA no fatty acid
  • BSA (no fatty acid) ( Sigma A-6003 ) 0.300 g Dissolve all dry powder ingredients except BSA in a beaker containing 70 ml of embryonic water. To prevent contamination of BME/MEM, add all liquid ingredients and cover the beaker with a cover. Stir, then add BSA dry powder and continue to stir until completely dissolved. The volume is adjusted to 100 ml, finally filtered through a 0.2 ⁇ m filter and stored at 4 Torr for no more than two weeks.
  • the nucleus of the first polar body of the recipient oocyte and its vicinity is removed by a denucleation needle with an inner diameter of about 20 ⁇ m, and then the donor nucleus of different mitochondrial DNA haplotypes is injected into the perivitelline space of the enucleated oocyte.
  • the nuclear nucleus was integrated into the oocyte at a fusion parameter of 2.5 Ky/cm - 3 V/cm and 6 - 10 ⁇ 3 to obtain a cloned reconstructed embryo.
  • the eggs after the fusion operation were transferred to an ACM culture solution, and cultured in an incubator at 5 % C0 2 , 38.5 ° C, and a saturated humidity. After 30 to 60 min, the fusion was examined under a stereo microscope, and the fusion rate was calculated. The results are shown in Table 3.
  • the above fused eggs were activated with ionomycin (Ionomycin, Sigma 1-0634) 5 ng/ml, and then placed in a culture dish containing CHX+CB (both purchased from sigma) and saturated at 38.5 °C. Place in a humidified incubator for 5 hours.
  • ionomycin Ionomycin, Sigma 1-0634
  • the reconstructed embryos were cultured in ACM medium and fetal rat fibroblasts (MEF) +1% fetal bovine serum (FBS), and cultured in an incubator at 5 % C0 2 , 38.5 ° C, and saturated humidity. After 72 h, the culture medium and MEF cells + 10% FBS were replaced. The cleavage condition of the reconstructed embryos was observed at 48h, the cleavage rate was calculated, and the number of blastocysts was recorded on the 7th day of culture, and the blastocyst rate was calculated. The results are shown in Table 3. Table 3. Comparison of different haplotype combinations and nuclear transfer efficiency between nucleoplasms
  • Fusion rate number of fusions / number of reconstructed embryos
  • Cleavage rate culture 48 hours split number / culture number
  • Blastocyst rate number of blastocysts/cleavage number obtained in 7 days of culture According to the results in Table 3, the nuclear transfer efficiency between the same mitochondrial DNA haplotype (homotype) and the difference between different mitochondrial DNA haplotypes (heterotype) were counted separately. The nuclear transfer efficiency, the results are shown in Table 4. Table 4. Comparison of different types of nuclear transfer efficiency
  • the fusion rate and cleavage rate of different combinations are basically the same, both above 70% and 54%; and in terms of blastocyst rate, the combination between the same haplotypes (A-A
  • the combination with C-C) has the best effect, all of which are above 40%, which is obviously better than the combination between different haplotypes (about 30%).
  • the blastocyst development rate of A-A combination was the highest, reaching 49.8 %; the C-C combination was second, reaching 42.2%; then the A ⁇ C combination was 36.1%, and the C-A combination development rate was the lowest, 24.4%. .
  • nuclear transfer using the same nucleus and cytoplasm of mitochondrial DNA haplotypes can effectively improve the efficiency of somatic cell nuclear transfer.
  • the use of the method of the present invention overcomes the problem of inefficient prior art nuclear transfer due to unclear oocyte origin.
  • the method of the present invention has been verified by taking the cattle as an example, those skilled in the art can clearly see that the method of the present invention is equally applicable to other non-human mammals, such as pigs, sheep, and mice, according to the contents of the specification. Therefore, a homologous nuclear transfer method for these non-human mammals should also fall within the scope of the present invention.

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  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
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  • General Engineering & Computer Science (AREA)
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  • Developmental Biology & Embryology (AREA)
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Abstract

L'invention concerne un procédé de transfert nucléaire de cellules, effectué entre une cellule somatique d'un donneur nucléaire mammifère non humain et un ovocyte receveur possédant le même haplotype d'ADN mitochondrial. Le procédé peut améliorer de façon remarquable la vitesse de fusion, la vitesse de clivage et la vitesse de formation de blastula des embryons reconstruits, et améliore par conséquent de façon remarquable l'efficacité du transfert nucléaire de cellules.
PCT/CN2007/002239 2006-08-03 2007-07-23 Procédé de transfert nucléaire de cellules WO2008017234A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN200610029674XA CN101117633B (zh) 2006-08-03 2006-08-03 一种细胞核移植方法
CN200610029674.X 2006-08-03

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WO2008017234A1 true WO2008017234A1 (fr) 2008-02-14

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Cited By (45)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015088643A1 (fr) 2013-12-11 2015-06-18 Regeneron Pharmaceuticals, Inc. Procédés et compositions pour la modification ciblée d'un génome
WO2015188109A1 (fr) 2014-06-06 2015-12-10 Regeneron Pharmaceuticals, Inc. Procédés et compositions pour la modification d'un locus ciblé
WO2015200805A2 (fr) 2014-06-26 2015-12-30 Regeneron Pharmaceuticals, Inc. Procédés et compositions pour modification génétiques ciblées et procédés d'utilisation
WO2016081923A2 (fr) 2014-11-21 2016-05-26 Regeneron Pharmaceuticals, Inc. Procédés et compositions pour modification génétique ciblée utilisant des arn guides appariés
WO2016196185A1 (fr) 2015-05-29 2016-12-08 Regeneron Pharmaceuticals, Inc. Animaux non-humains comprenant une perturbation dans un locus c9orf72
WO2017143062A1 (fr) 2016-02-16 2017-08-24 Regeneron Pharmaceuticals, Inc. Animaux non humains ayant un gène de kynuréninase mutant
WO2017201476A1 (fr) 2016-05-20 2017-11-23 Regeneron Pharmaceuticals, Inc. Procédés pour briser la tolérance immunologique à l'aide de multiples arn de guidage
WO2018023014A1 (fr) 2016-07-29 2018-02-01 Regeneron Pharmaceuticals, Inc. Souris comprenant des mutations entraînant l'expression de la fibrilline 1 tronquée en c
WO2018064600A1 (fr) 2016-09-30 2018-04-05 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant une expansion de répétition hexanucléotidique dans un locus c9orf72
WO2018157058A1 (fr) 2017-02-27 2018-08-30 Regeneron Pharmaceuticals, Inc. Modèles d'animaux non humains de rétinoschisis
WO2019006034A1 (fr) 2017-06-27 2019-01-03 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus asgr1 humanisé
WO2019028023A2 (fr) 2017-07-31 2019-02-07 Regeneron Pharmaceuticals, Inc. Procédés et compositions pour évaluer une disruption ou une excision médiée par crispr/cas et une recombinaison induite par crispr/cas à l'aide d'un acide nucléique donneur exogène in vivo
WO2019028029A1 (fr) 2017-07-31 2019-02-07 Regeneron Pharmaceuticals, Inc. Évaluation de la recombinaison induite par crispr/cas avec un acide nucléique donneur exogène in vivo
WO2019028032A1 (fr) 2017-07-31 2019-02-07 Regeneron Pharmaceuticals, Inc. Cellules souches embryonnaires de souris transgéniques cas et souris et leurs utilisations
EP3456831A1 (fr) 2013-04-16 2019-03-20 Regeneron Pharmaceuticals, Inc. Modification ciblée du génome d'un rat
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WO2019067875A1 (fr) 2017-09-29 2019-04-04 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus ttr humanisé et procédés d'utilisation
US10285387B2 (en) 2015-03-16 2019-05-14 Regeneron Pharmaceuticals, Inc. Non-human animal exhibiting diminished upper and lower motor neuron function and sensory perception
WO2019094735A1 (fr) 2017-11-10 2019-05-16 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant une mutation de slc30a8 et procédés d'utilisation
WO2019108983A1 (fr) 2017-11-30 2019-06-06 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus trkb humanisé
WO2019183123A1 (fr) 2018-03-19 2019-09-26 Regeneron Pharmaceuticals, Inc. Modulation de la transcription chez des animaux à l'aide de systèmes crispr/cas
WO2020056122A1 (fr) 2018-09-13 2020-03-19 Regeneron Pharmaceuticals, Inc. Rat à invalidation génique de facteur h de complément en tant que modèle de glomérulopathie c3
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WO2020131632A1 (fr) 2018-12-20 2020-06-25 Regeneron Pharmaceuticals, Inc. Expansion de répétition à médiation par nucléase
WO2020206139A1 (fr) 2019-04-04 2020-10-08 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus facteur 12 de coagulation humanisé
WO2020247812A1 (fr) 2019-06-07 2020-12-10 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus d'albumine humanisé
WO2020247452A1 (fr) 2019-06-04 2020-12-10 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus ttr humanisé ayant une mutation bêta-slip et procédés d'utilisation
WO2020264339A1 (fr) 2019-06-27 2020-12-30 Regeneron Pharmaceuticals, Inc. Modélisation de la protéinopathie à tdp-43
WO2021108363A1 (fr) 2019-11-25 2021-06-03 Regeneron Pharmaceuticals, Inc. Régulation à la hausse médiée par crispr/cas d'un allèle ttr humanisé
WO2021154791A1 (fr) 2020-01-28 2021-08-05 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus pnpla3 humanisé et procédés d'utilisation
WO2021158883A1 (fr) 2020-02-07 2021-08-12 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus klkb1 humanisé et procédés d'utilisation
WO2021195079A1 (fr) 2020-03-23 2021-09-30 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus ttr humanisé affichant une mutation v30m et méthodes d'utilisation
WO2021263146A2 (fr) 2020-06-26 2021-12-30 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus d'ace2 humanisé
WO2023081847A1 (fr) 2021-11-04 2023-05-11 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus cacng1 modifié
WO2023108047A1 (fr) 2021-12-08 2023-06-15 Regeneron Pharmaceuticals, Inc. Modèle de maladie impliquant une myociline mutante et ses utilisations
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WO2023150798A1 (fr) 2022-02-07 2023-08-10 Regeneron Pharmaceuticals, Inc. Compositions et procédés permettant de définir des délais de traitement optimaux dans le cadre d'une maladie lysosomale
WO2023154861A1 (fr) 2022-02-11 2023-08-17 Regeneron Pharmaceuticals, Inc. Compositions et procédés de criblage d'agents de ciblage de tau 4r
WO2023235677A1 (fr) 2022-05-31 2023-12-07 Regeneron Pharmaceuticals, Inc. Modèle animal de protéinopathie tdp-43
WO2024026488A2 (fr) 2022-07-29 2024-02-01 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus de récepteur de transférrine modifié
WO2024031053A1 (fr) 2022-08-05 2024-02-08 Regeneron Pharmaceuticals, Inc. Variants de tdp-43 résistants à l'agrégation
WO2024073679A1 (fr) 2022-09-29 2024-04-04 Regeneron Pharmaceuticals, Inc. Correction de la stéatose hépatique chez des animaux hépatiques humanisés par restauration de la signalisation il6/il6r/gp130 dans des hépatocytes humains
WO2024163650A1 (fr) 2023-02-01 2024-08-08 Regeneron Pharmaceuticals, Inc. Animaux comprenant un locus klhdc7b modifié
WO2025006963A1 (fr) 2023-06-30 2025-01-02 Regeneron Pharmaceuticals, Inc. Procédés et compositions pour augmenter la réparation dirigée par homologie
WO2025122754A1 (fr) 2023-12-07 2025-06-12 Regeneron Pharmaceuticals, Inc. Animaux non humains knockout pour gaa

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101665780B (zh) * 2008-09-04 2011-06-08 中国科学院动物研究所 一种提高体细胞重编程效率的方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020155601A1 (en) * 2001-01-02 2002-10-24 Yan Wen Liang Method for producing a population of homozygous stem cells having a pre-selected immunotype and/or genotype, cells suitable for transplant derived therefrom, and materials and methods using same
WO2003100018A2 (fr) * 2002-05-24 2003-12-04 Advanced Cell Technology, Inc. Banque de cellules souches destinees a la production de cellules pour transplantation possedant des antigenes hla correspondant a ceux des receveurs de transplant, et procedes de constitution et d'utilisation d'une telle banque de cellules souches
CN1557946A (zh) * 2004-02-11 2004-12-29 上海市儿童医院 一种同体细胞核移植方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020155601A1 (en) * 2001-01-02 2002-10-24 Yan Wen Liang Method for producing a population of homozygous stem cells having a pre-selected immunotype and/or genotype, cells suitable for transplant derived therefrom, and materials and methods using same
WO2003100018A2 (fr) * 2002-05-24 2003-12-04 Advanced Cell Technology, Inc. Banque de cellules souches destinees a la production de cellules pour transplantation possedant des antigenes hla correspondant a ceux des receveurs de transplant, et procedes de constitution et d'utilisation d'une telle banque de cellules souches
CN1557946A (zh) * 2004-02-11 2004-12-29 上海市儿童医院 一种同体细胞核移植方法

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
"Effect of Oocyte Mitochondrial DNA Haplotype on Bovine Somatic Cell Nuclear Transfer Efficiency", MOLECULAR REPRODUCTION AND DEVELOPMENT, vol. 74, 8 February 2007 (2007-02-08), pages 1278 - 1286 *
HIENDLEDER S. AND WOLF E.: "The mitochondrial genome in embryo technologies", REPRODUCTION IN DOMESTIC ANIMALS, vol. 38, no. 4, 31 October 2003 (2003-10-31), pages 290 - 304 *
JIAO F.: "Identification of Genetic Materials in Clonsed Cattle by Somatic Cell Nuclear Transfer", JOURNAL OF THE SHANGHAI JIAOTONG UNIVERSITY (AGRICULTURAL SCIENCE), vol. 24, no. 3, 30 June 2006 (2006-06-30) *
STEINBORN R. ET AL.: "Coexistence of Bos taurus and B. indicus Mitochondrial DNAs in Nuclear Transfer-Derived Somatic Cattle Clones", GENETICS, vol. 162, 31 October 2002 (2002-10-31), pages 823 - 829, XP002969934 *

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US11547101B2 (en) 2015-05-29 2023-01-10 Regeneron Pharmaceuticals, Inc. Non-human animals having a disruption in a C9ORF72 locus
EP3689139A1 (fr) 2015-05-29 2020-08-05 Regeneron Pharmaceuticals, Inc. Animaux non-humains comprenant une perturbation dans un locus c9orf72
US10285388B2 (en) 2015-05-29 2019-05-14 Regeneron Pharmaceuticals, Inc. Non-human animals having a disruption in a C9ORF72 locus
WO2016196185A1 (fr) 2015-05-29 2016-12-08 Regeneron Pharmaceuticals, Inc. Animaux non-humains comprenant une perturbation dans un locus c9orf72
WO2017143062A1 (fr) 2016-02-16 2017-08-24 Regeneron Pharmaceuticals, Inc. Animaux non humains ayant un gène de kynuréninase mutant
EP4368637A2 (fr) 2016-05-20 2024-05-15 Regeneron Pharmaceuticals, Inc. Procédés pour briser la tolérance immunologique à l'aide de multiples arn de guidage
WO2017201476A1 (fr) 2016-05-20 2017-11-23 Regeneron Pharmaceuticals, Inc. Procédés pour briser la tolérance immunologique à l'aide de multiples arn de guidage
WO2018023014A1 (fr) 2016-07-29 2018-02-01 Regeneron Pharmaceuticals, Inc. Souris comprenant des mutations entraînant l'expression de la fibrilline 1 tronquée en c
US10781453B2 (en) 2016-09-30 2020-09-22 Regeneron Pharmaceuticals, Inc. Non-human animals having a hexanucleotide repeat expansion in a C9ORF72 locus
WO2018064600A1 (fr) 2016-09-30 2018-04-05 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant une expansion de répétition hexanucléotidique dans un locus c9orf72
WO2018157058A1 (fr) 2017-02-27 2018-08-30 Regeneron Pharmaceuticals, Inc. Modèles d'animaux non humains de rétinoschisis
WO2019006034A1 (fr) 2017-06-27 2019-01-03 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus asgr1 humanisé
US11696572B2 (en) 2017-06-27 2023-07-11 Regeneron Pharmaceuticals, Inc. Non-human animals comprising a humanized ASGR1 locus
EP4484562A2 (fr) 2017-06-27 2025-01-01 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus asgr1 humanisé
WO2019028023A2 (fr) 2017-07-31 2019-02-07 Regeneron Pharmaceuticals, Inc. Procédés et compositions pour évaluer une disruption ou une excision médiée par crispr/cas et une recombinaison induite par crispr/cas à l'aide d'un acide nucléique donneur exogène in vivo
WO2019028032A1 (fr) 2017-07-31 2019-02-07 Regeneron Pharmaceuticals, Inc. Cellules souches embryonnaires de souris transgéniques cas et souris et leurs utilisations
WO2019028029A1 (fr) 2017-07-31 2019-02-07 Regeneron Pharmaceuticals, Inc. Évaluation de la recombinaison induite par crispr/cas avec un acide nucléique donneur exogène in vivo
WO2019067875A1 (fr) 2017-09-29 2019-04-04 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus ttr humanisé et procédés d'utilisation
EP4276185A2 (fr) 2017-09-29 2023-11-15 Regeneron Pharmaceuticals, Inc. Rongeurs comprenant un locus ttr humanisé et procédés d'utilisation
WO2019094735A1 (fr) 2017-11-10 2019-05-16 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant une mutation de slc30a8 et procédés d'utilisation
WO2019108983A1 (fr) 2017-11-30 2019-06-06 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus trkb humanisé
EP4299732A2 (fr) 2017-11-30 2024-01-03 Regeneron Pharmaceuticals, Inc. Rats comprenant un locus trkb humanisé
WO2019183123A1 (fr) 2018-03-19 2019-09-26 Regeneron Pharmaceuticals, Inc. Modulation de la transcription chez des animaux à l'aide de systèmes crispr/cas
WO2020056122A1 (fr) 2018-09-13 2020-03-19 Regeneron Pharmaceuticals, Inc. Rat à invalidation génique de facteur h de complément en tant que modèle de glomérulopathie c3
WO2020131632A1 (fr) 2018-12-20 2020-06-25 Regeneron Pharmaceuticals, Inc. Expansion de répétition à médiation par nucléase
US11690362B2 (en) 2018-12-20 2023-07-04 Regeneran Pharmaceuticals, Inc. Nuclease-mediated repeat expansion
WO2020206139A1 (fr) 2019-04-04 2020-10-08 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus facteur 12 de coagulation humanisé
WO2020247452A1 (fr) 2019-06-04 2020-12-10 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus ttr humanisé ayant une mutation bêta-slip et procédés d'utilisation
WO2020247812A1 (fr) 2019-06-07 2020-12-10 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus d'albumine humanisé
WO2020264339A1 (fr) 2019-06-27 2020-12-30 Regeneron Pharmaceuticals, Inc. Modélisation de la protéinopathie à tdp-43
WO2021108363A1 (fr) 2019-11-25 2021-06-03 Regeneron Pharmaceuticals, Inc. Régulation à la hausse médiée par crispr/cas d'un allèle ttr humanisé
WO2021154791A1 (fr) 2020-01-28 2021-08-05 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus pnpla3 humanisé et procédés d'utilisation
WO2021158883A1 (fr) 2020-02-07 2021-08-12 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus klkb1 humanisé et procédés d'utilisation
WO2021195079A1 (fr) 2020-03-23 2021-09-30 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus ttr humanisé affichant une mutation v30m et méthodes d'utilisation
WO2021263146A2 (fr) 2020-06-26 2021-12-30 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus d'ace2 humanisé
WO2023081847A1 (fr) 2021-11-04 2023-05-11 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus cacng1 modifié
WO2023108047A1 (fr) 2021-12-08 2023-06-15 Regeneron Pharmaceuticals, Inc. Modèle de maladie impliquant une myociline mutante et ses utilisations
WO2023122506A1 (fr) 2021-12-20 2023-06-29 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant des loci ace2 et tmprss humanisés
WO2023150798A1 (fr) 2022-02-07 2023-08-10 Regeneron Pharmaceuticals, Inc. Compositions et procédés permettant de définir des délais de traitement optimaux dans le cadre d'une maladie lysosomale
WO2023154861A1 (fr) 2022-02-11 2023-08-17 Regeneron Pharmaceuticals, Inc. Compositions et procédés de criblage d'agents de ciblage de tau 4r
WO2023235677A1 (fr) 2022-05-31 2023-12-07 Regeneron Pharmaceuticals, Inc. Modèle animal de protéinopathie tdp-43
WO2024026488A2 (fr) 2022-07-29 2024-02-01 Regeneron Pharmaceuticals, Inc. Animaux non humains comprenant un locus de récepteur de transférrine modifié
WO2024031053A1 (fr) 2022-08-05 2024-02-08 Regeneron Pharmaceuticals, Inc. Variants de tdp-43 résistants à l'agrégation
WO2024073679A1 (fr) 2022-09-29 2024-04-04 Regeneron Pharmaceuticals, Inc. Correction de la stéatose hépatique chez des animaux hépatiques humanisés par restauration de la signalisation il6/il6r/gp130 dans des hépatocytes humains
WO2024163650A1 (fr) 2023-02-01 2024-08-08 Regeneron Pharmaceuticals, Inc. Animaux comprenant un locus klhdc7b modifié
WO2025006963A1 (fr) 2023-06-30 2025-01-02 Regeneron Pharmaceuticals, Inc. Procédés et compositions pour augmenter la réparation dirigée par homologie
WO2025122754A1 (fr) 2023-12-07 2025-06-12 Regeneron Pharmaceuticals, Inc. Animaux non humains knockout pour gaa

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