Generating a transgenic or gene-modified mouse requires the introduction of exogenous reagents into an early-stage embryo. The mouse one-cell embryo or zygote possesses two pronuclei, representing the genetic contribution of the sperm and oocyte. Traditional transgenic mice are generated by injecting a DNA solution containing a purified transgene construct into the male pronucleus, generally the larger of the two pronuclei. Similarly, gene-editing reagents such as ZFNs, TALENs, and CRISPR RNAs are introduced into zygotes in the same manner, making this technique applicable to a wide variety of projects. This chapter presents the procedures for pronuclear microinjection.
CITATION STYLE
Larson, M. A. (2020). Pronuclear Microinjection of One-Cell Embryos. In Methods in Molecular Biology (Vol. 2066, pp. 27–33). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9837-1_2
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