PMID: 7581516Nov 1, 1995Paper

Transgenesis in mice by cytoplasmic injection of polylysine/DNA mixtures

Transgenic Research
R L PageW H Velander

Abstract

Pronuclear injection is currently the most often used method to make transgenic animals, but in some animal species it is temporally restrictive due to difficulty in visualizing pronuclei. However, the injection of construct DNA into the cytoplasm does not result in transgenesis. The production of transgenic mice by a cytoplasmic microinjection technique of polylysine complexed DNA into pronuclear stage zygotes is described. Transgenic mice were produced from cytoplasmic microinjection of mixtures of a 5.3 kb linearized DNA and poly-L-lysine (degree of polymerization = 51). Effects on transgenic frequency of both the lysine to phosphate ratio of polylysine to DNA and DNA concentration were studied. About 12.8% of the pups born from zygotes cytoplasmically microinjected with a polylysine/DNA mixture having a lysine to phosphate ratio (L:P) of 1:1 at a DNA concentration of 50 micrograms ml-1 were transgenic. The transgenic frequency for the pronuclear microinjection positive control of DNA alone was 21.7%. No transgenic pups were born from microinjection of DNA alone into the cytoplasm. Complexes of polylysine/DNA were detected using agarose gel electrophoresis at the conditions which produced transgenic mice. The presence of pol...Continue Reading

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Citations

Sep 2, 1999·Molecular Reproduction and Development·M M IvanovaL K Ernst
Mar 28, 2013·Transgenic Research·Aubrey M WilsonSandra H Burnett
Sep 29, 2011·Medical & Biological Engineering & Computing·Hadi EsmaeilsabzaliEdward J Park
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Sep 3, 2002·Advanced Drug Delivery Reviews·Thomas MerdanThomas Kissel
Jun 6, 2000·Advanced Drug Delivery Reviews·E WagnerW Zauner
Jun 1, 2010·Zygote : the Biology of Gametes and Early Embryos·P ChrenekA V Makarevich
Mar 4, 2006·Reproduction : the Official Journal of the Society for the Study of Fertility·Augusta ZannoniMonica Forni
May 20, 1999·Molecular Membrane Biology·N MaurerP R Cullis
Nov 22, 2005·Expert Opinion on Drug Delivery·Sérgio SimõesMaria Pedroso de Lima
Jun 5, 2016·Reproduction in Domestic Animals = Zuchthygiene·C G LuchettiD F Salamone
Nov 17, 2007·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·H EliyahuA J Domb
Dec 14, 2012·PloS One·Age BrauerMaido Remm
Oct 27, 2015·Journal of Animal Science and Biotechnology·Fanli MengDeshun Shi

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