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Maternal mRNA Knock-down Studies: Antisense Experiments Using the Host-Transfer Technique in  Xenopus laevis and  Xenopus tropicalis
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Maternal mRNA Knock-down Studies: Antisense Experiments Using the Host-Transfer Technique in Xenopus laevis and Xenopus tropicalis

David J Olson, Alissa M Hulstrand and Douglas W Houston
Xenopus Protocols: Post-Genomic Approaches, Vol.917, pp.167-182
Methods in Molecular Biology, 917, 2
2012
DOI: 10.1007/978-1-61779-992-1_10
PMCID: PMC3974620
PMID: 22956088
url
https://doi.org/10.1007/978-1-61779-992-1_10View
Published (Version of record) Open Access

Abstract

The ability to inhibit the activity of maternally stored gene products in Xenopus has led to numerous insights into early developmental mechanisms. Oocytes can be cultured and manipulated in vitro and then implanted into the body cavity of a host female to make them competent for fertilization. Here, we summarize the methods for obtaining, culturing, and fertilizing Xenopus oocytes, with the goal of inhibiting maternal gene function through antisense oligonucleotide-mediated mRNA knock-down. We describe a simplified technique for implanting donor oocytes into host females using intraperitoneal injection. Also, we present optimized methods for performing the host-transfer procedure with Xenopus tropicalis oocytes.
oocyte host-transfer Xenopus laevis maternal genes oligonucleotides Xenopus tropicalis antisense

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