Cite as: Cold Spring Harb. Protoc.; 2009; doi:10.1101/pdb.prot5347

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Microinjection of Morpholino Oligos and RNAs in Sea Squirt (Ciona) Embryos

Lionel Christiaen1,2, Eileen Wagner, Weiyang Shi, and Michael Levine2

Molecular and Cell Biology Department, University of California, Berkeley, California 94720, USA

1 Present address: Center for Developmental Genetics, Department of Biology, Faculty of Arts and Science, New York University, New York, NY 10003, USA.

2Corresponding authors (mlevine{at}berkeley.edu); (lc121{at}nyu.edu).


INTRODUCTION

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 of pleiotropic genes. Injection can also be performed in unfertilized eggs to inhibit maternal genes. Microinjection also permits DiI labeling and lineage tracing.


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This article has been cited by other articles:


Home page
 Cold Spring Harb ProtocHome page
L. Christiaen, E. Wagner, W. Shi, and M. Levine
The Sea Squirt Ciona intestinalis
Cold Spring Harb Protoc, December 1, 2009; 2009(12): 10.1101/pdb.emo138.
[Abstract] [Full Text]

Home page
 Cold Spring Harb ProtocHome page
L. Christiaen, E. Wagner, W. Shi, and M. Levine
Isolation of Sea Squirt (Ciona) Gametes, Fertilization, Dechorionation, and Development
Cold Spring Harb Protoc, December 1, 2009; 2009(12): 10.1101/pdb.prot5344.
[Abstract] [Full Text]