Microinjection of Antisense Morpholinos, CRISPR/Cas9 RNP, and RNA/DNA into Zebrafish Embryos

Yi Xin; Cunming Duan

doi:10.1007/978-1-4939-7665-2_18

Microinjection of Antisense Morpholinos, CRISPR/Cas9 RNP, and RNA/DNA into Zebrafish Embryos

Methods Mol Biol. 2018:1742:205-211. doi: 10.1007/978-1-4939-7665-2_18.

Authors

Yi Xin¹, Cunming Duan²

Affiliations

¹ Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, USA.
² Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, USA. cduan@umich.edu.

PMID: 29330802
DOI: 10.1007/978-1-4939-7665-2_18

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.

Keywords: CRISPR/Cas9; Gene knockdown; Microinjection; Morpholinos; Transgenesis; Zebrafish embryos; mRNA.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
CRISPR-Cas Systems
Embryonic Development
Female
Gene Expression Regulation, Developmental
Genetic Engineering / instrumentation*
Microinjections / methods*
Models, Animal
Morpholinos / administration & dosage
Oligonucleotides, Antisense / administration & dosage
RNA, Guide, CRISPR-Cas Systems / administration & dosage
Ribonucleoproteins / administration & dosage
Thionucleotides
Zebrafish / embryology*
Zebrafish / genetics
Zebrafish / growth & development
Zygote / growth & development

Substances

Morpholinos
Oligonucleotides, Antisense
RNA, Guide, CRISPR-Cas Systems
Ribonucleoproteins
Thionucleotides