Microinjection of morpholino oligos and RNAs in sea squirt (Ciona) embryos

Lionel Christiaen, Eileen Wagner, Weiyang Shi, Michael Levine

Research output: Contribution to journalArticle

Abstract

This protocol describes microinjection of morpholino oligos (MOs) and RNAs into sea squirt (Ciona) embryos. This is the method of choice for gene disruption assays. MOs that target the initiating ATG can be used, in addition to those that target splice donor and acceptor sites. The latter method permits the selective inhibition of zygotic mRNAs in cases in which the gene in question is expressed in both the egg and embryo. Although injection is usually performed at the one-cell stage, it is possible to target individual blastomeres, up to the eight-cell stage, thereby permitting lineage-specific knockdown ofpleiotropic genes. Injection can also be performed in unfertilized eggs to inhibit maternal genes. Microinjection also permits DiI labeling and lineage tracing.

Original languageEnglish (US)
JournalCold Spring Harbor Protocols
Volume4
Issue number12
DOIs
StatePublished - 2009

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Morpholinos
Microinjections
Oceans and Seas
RNA Splice Sites
Embryonic Structures
Genes
RNA
Ovum
Gene Knockdown Techniques
Blastomeres
Injections
Mothers
Labeling
Assays
Messenger RNA

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Microinjection of morpholino oligos and RNAs in sea squirt (Ciona) embryos. / Christiaen, Lionel; Wagner, Eileen; Shi, Weiyang; Levine, Michael.

In: Cold Spring Harbor Protocols, Vol. 4, No. 12, 2009.

Research output: Contribution to journalArticle

Christiaen, Lionel ; Wagner, Eileen ; Shi, Weiyang ; Levine, Michael. / Microinjection of morpholino oligos and RNAs in sea squirt (Ciona) embryos. In: Cold Spring Harbor Protocols. 2009 ; Vol. 4, No. 12.
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