Biopolym. Cell. 2021; 37(4):289-302.
http://dx.doi.org/10.7124/bc.000A5B
Molecular and Cell Biotechnologies
Efficacy of blastodermal cells and CRISPR/CAS9 method in the creation of transgenic duck (Anas Platyrhynchos)
1, 2Konoval O., 3Korol P., 4Kostenko S., 1Tabaka P., 1Lizhi L., 4Chepiha A., 4Doroshenko M., 3Sydorenko O., 3Dzhus P., 4Svyrydenko N., 4Lytvynenko T., 1Xingchen B., 5Xuetao H., 5Li L., 6Drahulian M., 4Kostyuk E., 4Filipova P.
  1. Institute of Animal Husbandry and Veterinary Science, Zhejiang Academy of Agricultural Sciences
    198 Shiqiao Road, Hangzhou, Zhejiang, China, 310021
  2. Ukrainian Laboratory of Quality and Safety of Agricultural Products
    National University of Life and Environmental Sciences of Ukraine
    7, Mashynobudivnykiv Str., Chabany, Kyiv region, Ukraine, 08162
  3. Institute of Animal Breeding and Genetics nd. a. M.V. Zubets of NAAS of Ukraine,
    1, Pohrebniaka Str., Chubyns’ke, Kyivs’ka obl., Ukraine, 083213
  4. National University of Life and Environmental Sciences of Ukraine
    15, Heroiv Oborony St., Kyiv, Ukraine,03041
  5. Zhuji Guowey Poultry Development Co, Ltd, Ltd.
    1, Gujing Road, Wangjiajing, Zhuji, Zhejiang, China, 311813
  6. Rinderunion Baden-Württemberg Genetik GmbH,
    Hopfenweilerweg 20,88339 Bad Waldsee, Germany

Abstract

Aim To develop a methodology for the creation of transgenic chimeras of ducks by using donor blastodermal cells after transfection with DNA vector and Lipofectamine 2000® (Invitrogen, USA). Methods. Transgenic duck chimeras were created using donor blastodermal cells after transfection with plasmid DNA and Lipofectamine 2000. To edit the target region of the duck genome, we used the CRISPR / cas9 system with HDR. The EGFP reporter gene was used as the transgene. Results. Of the 200 eggs, in which the transfected blastodermal cells were introduced, 20 offspring were obtained, including 8 males and 12 females. Thus, the survival of embryos was 10 %. Among the surviving fertile animals, 13/20 were animals G0 (65 %): 10/12 (83.3 %) females and 3/8 (37.5 %) males. The procedure of obtaining chimeras has a stronger effect on the survival and fertility of male chimeras. From 13 of 20 birds G0, we received a total of 197 offspring (including 117 (59.4 %) daughters and 80 (40.6 %) sons), 59 of which were EGFP-positive (30.3 %), including 10 males (16.9 %) and 49 females (83.1 %). Conclusions. The technique used by us can be successfully applied in further researches and for creation of a transgenic duck.
Keywords: CRISPR/Cas9, EGFP, transgenic duck, blastodermal bells, chimera
Full text: (PDF, in English)

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