Document Detail


Morpholino injection in Xenopus.
MedLine Citation:
PMID:  22222519     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The study of gene function in developmental biology has been significantly furthered by advances in antisense technology made in the early 2000s. This was achieved, in particular, by the introduction of morpholino (MO) oligonucleotides. The introduction of antisense MO oligonucleotides into cells enables researchers to readily reduce the levels of their protein of interest without investing huge financial or temporal resources, in both in vivo and in vitro model systems. Historically, the African clawed frog Xenopus has been used to study vertebrate embryological development, due to its ability to produce vast numbers of offspring that develop rapidly, in synchrony, and can be cultured in buffers with ease. The developmental progress of Xenopus embryos has been extensively characterized and this model organism is very easy to maintain. It is these attributes that enable MO-based knockdown strategies to be so effective in Xenopus. In this chapter, we will detail the methods of microinjecting MO oligonucleotides into early embryos of X. laevis and X. tropicalis. We will discuss how MOs can be used to prevent either pre-mRNA splicing or translation of the specific gene of interest resulting in abrogation of that gene's function and advise on what control experiments should be undertaken to verify their efficacy.
Authors:
Panna Tandon; Chris Showell; Kathleen Christine; Frank L Conlon
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  843     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2012  
Date Detail:
Created Date:  2012-01-06     Completed Date:  2012-04-26     Revised Date:  2014-10-28    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  29-46     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Western
Cell Extracts
Embryo, Nonmammalian / cytology,  metabolism
Gene Knockdown Techniques
Humans
Microinjections / instrumentation,  methods*
Morpholinos / administration & dosage*,  genetics*
Protein Biosynthesis / genetics
Reverse Transcriptase Polymerase Chain Reaction
Xenopus laevis / embryology*,  metabolism*
Grant Support
ID/Acronym/Agency:
R01 DE018825/DE/NIDCR NIH HHS; T32 HL069768/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Cell Extracts; 0/Morpholinos
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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