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24th Biotechnology Congress: Research & Innovations
CRISPR Cas9 Technology and Genetic Engineering
Annual Congress on
October 24-25, 2018 | Boston, USA
Journal of Biotechnology & Biomaterials | ISSN: 2155-952X | Volume: 8
Generation of transgenic chimeric ducks using blastoderm cell transfer CRISPR/Cas9-mediated
gene insertion into the duck genome
Oksana Konoval 1 3, Maria Doroshenko2, Svitlana Kostenko 1,2,5, Lizhi Lu1, Pavlo Tabaka1,3, Petro Korol4, Alona Chepiha2, Xingchen Bu 1, Lingling Huang3,
Xuetao Huang5, Liumeng Li5
1Zhejiang Academy of Agricultural Sciences(Hangzhou, China)
2National University of Life and Environmental Sciences of Ukraine (Kiev, Ukraine)
3Ukrainian Laboratory of Quality and Safety of Agricultural Products of National University of Life and Environmental Sciences of Ukraine(Chabany, Ukraine)
4Institute of Animal Breeding and Genetics nd. a. M.V. Zubets NAAS of Ukraine (Tchoubynske, Ukraine)
5Zhuji Guowey Poultry Development Co, Ltd (Zhuji, China)
Statement of the Problem: The ability to culture and genetically modify embryonic cells changed developmental biology. The
production of transgenic birds has increasing applications in biotechnology. There are various methods of birds transgenesis
and the production of germline chimeras. The technique of injection DNA under the germinal disk could be used as one of
the effective and often used methods. Weak reproductive ability and low survival rate of the produced chimeras are the main
disadvantages of this method.
Methodology & Theoretical Orientation: In order to produce germline chimeras, the embryos of the Shan partridge duck
were used as recipients, and blastodermal embryos of Shaoxing ducks were used as donors. Recipients were sterilized using
ultraviolet light irradiation. The isolated blastodermal donor cells were transfected with the DNA-construction (CRISPR/
Cas9-mediated gene insertion into the duck genome) with lipofectamine which was inoculated under embryo of the of the
recipient eggs.
Findings: Survival rate of recipient embryos following transfected was 6.98% (19/272). After hatching, eleven female and eight
male alive birds (3 % of the manipulated embryos) were obtained. Five of eleven female founders and five of eight male founders
carried the transgene construct, and the transgenic bird production efficiency was 44.4% (from survive).Among the five male
founders, only one (number 28) was most fertile. It produced 34 descendants (18 females and 14 males), of which two sons
and nine daughters were transgenic (32.2%).Thus, the obtained data indicates that this technique the positive outlook on usage
CRISPR/Cas9-mediated gene insertion into the duck genome of the Shan partridge and Shaoxing duck model for creating
transgenic ducks and should be useful in developmental studies and may facilitate the production of transgenic poultry as the
exogenous DNA was successfully inserted into the duck genome.
This study was supported by the Earmarked Fund for National Waterfowl-industry Technology Research System ( CARS-42-06)
and the Zhejiang Major Scientific and Technological Project of Agricultural (livestock's) Breeding (grant number 2016C02054-12).
Biography
Mariia Doroshenko is a doctor of veterinary medicine and a PhD student at the Department of Genetics, Breeding and Biotechnology of Animals at the National
University of Life and Environmental Sciences of Ukraine. Her work is based on gene polymorphism in breeding. She has engaged in genetics and breeding in
poultry work for more than two years.
Oksana Konoval1 3 et al., J Biotechnol Biomater 2018, Volume 8
DOI: 10.4172/2155-952X-C4-098