Abstract
In this chapter, we describe a stepwise protocol of microinjection. Using this method, antisense morpholinos, CRISPR-Cas9 ribonucleoprotein complexes, capped mRNA, and DNA can be delivered into fertilized zebrafish eggs to manipulate gene expression during development. This protocol can also be adapted for microinjection in other fish and amphibian species.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Hogan BM, Verkade H, Lieschke GJ, Heath JK (2008) Manipulation of gene expression during zebrafish embryonic development using transient approaches. Methods Mol Biol 469:273–300
Suster ML, Abe G, Schouw A, Kawakami K (2011) Transposon-mediated BAC transgenesis in zebrafish. Nat Protoc 6:1998–2021
Burger A, Lindsay H, Felker A (2016) Maximizing mutagenesis with solubilized CRISPR-Cas9 ribonucleoprotein complexes. Development 143:2025–2037
Robu ME, Larson JD, Nasevicius A, Beiraghi S, Brenner C, Farber SA, Ekker SC (2007) p53 activation by knockdown technologies. PLoS Genet 3(5):e78
Westerfield M (2007) The zebrafish book. A guide for the laboratory use of zebrafish (Danio rerio), 5th edn. University of Oregon Press, Eugene
Kimmel CB, Ballard WW, Kimmel SR, Ullmann B, Schilling TF (1995) Stages of embryonic development of the zebrafish. Dev Dyn 203(3):253–310
Brocal I, White RJ, Dooley CM, Carruthers SN, Clark R, Hall A, Busch-Nentwich EM, Stemple DL, Kettleborough RN (2016) Efficient identification of CRISPR/Cas9-induced insertions/deletions by direct germline screening in zebrafish. BMC Genomics 17:259
Acknowledgments
We thank Ms. Allison Malick, University of Michigan, for proofreading this manuscript. This work was supported by National Science Foundation grant IOS-1557850 and University of Michigan MCube2.0 Project U0496246 to CD.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Xin, Y., Duan, C. (2018). Microinjection of Antisense Morpholinos, CRISPR/Cas9 RNP, and RNA/DNA into Zebrafish Embryos. In: Huang, L. (eds) Hypoxia. Methods in Molecular Biology, vol 1742. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7665-2_18
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7665-2_18
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7664-5
Online ISBN: 978-1-4939-7665-2
eBook Packages: Springer Protocols