Suppressing mosaicism by Au nanowire injector-driven direct delivery of plasmids into mouse embryos

Transgenic animals have become key tools in a variety of biomedical research areas. However, microinjection commonly used for producing transgenic animals has several challenges such as physical and chemical damage to the embryos due to microinjector with buffer, and low transgene integration rates with frequent mosaicism. Here, we report direct delivery of plasmids into mouse embryos using a Au nanowire injector (NWI) that significantly improved transgene integration efficiency and suppressed mosaicism. The Au NWI could deliver plasmid into the pronucleus (PN) of a mouse zygote without buffer and rapidly release it with electric pulse. Because zygote, which is a fertilized 1-cell stage embryo, has two physical barriers (cytoplasmic membrane and zona pellucida), direct delivery of plasmids into PN of zygote is more difficult than into a normal cell type. To penetrate the two physical barriers with minimal disruption of the embryo, we optimized the diameter and length of Au NWI. The mosaicism is more reduced in the Au NWI injected embryos than in micropipette injected embryos, which was determined by the expression of transgene in a blastocyst stage. We suggest that Au NWI can increase the efficiency of gene delivery into zygote with suppressed mosaicism and become a useful alternative. (C) 2017 Published by Elsevier Ltd.
Publisher
ELSEVIER SCI LTD
Issue Date
2017-09
Language
English
Keywords

GERM-LINE TRANSMISSION; TRANSGENIC MICE; SILICON NANOWIRES; MAMMALIAN-CELLS; GENE DELIVERY; CARBON NANOTUBES; LIVING CELLS; STEM-CELLS; DNA; EFFICIENCY

Citation

BIOMATERIALS, v.138, pp.169 - 178

ISSN
0142-9612
DOI
10.1016/j.biomaterials.2017.05.044
URI
http://hdl.handle.net/10203/224702
Appears in Collection
CH-Journal Papers(저널논문)
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