Advertisement

Pronuclear Microinjection of One-Cell Embryos

  • Melissa A. LarsonEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2066)

Abstract

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.

Key words

Transgenic CRISPR Microinjection Zygotes In vivo Gene editing 

References

  1. 1.
    Gordon JW, Scangos GA, Plotkin DJ, Barbosa JA, Ruddle FH (1980) Genetic transformation of mouse embryos by microinjection of purified DNA. Proc Natl Acad Sci U S A 77:7380–7384CrossRefGoogle Scholar
  2. 2.
    Gordon JW, Ruddle FH (1981) Integration and stable germ line transmission of genes injected into mouse pronuclei. Science 214(4526):1244–1246CrossRefGoogle Scholar
  3. 3.
    Brinster RL, Chen HY, Trumbauer ME, Yagle MK, Palmiter RD (1985) Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs. Proc Natl Acad Sci U S A 82:4438–4442CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Transgenic and Gene-Targeting FacilityUniversity of Kansas Medical CenterKansas CityUSA
  2. 2.Department of Molecular and Integrative PhysiologyUniversity of Kansas Medical CenterKansas CityUSA

Personalised recommendations