Defining synphenotype groups in Xenopus tropicalis by use of antisense morpholino oligonucleotides.
To identify novel genes involved in early development, and as proof-of-principle of a large-scale reverse genetics approach in a vertebrate embryo, we have carried out an antisense morpholino oligonucleotide (MO) screen in Xenopus tropicalis, in the course of which we have targeted 202 genes expressed during gastrula stages. MOs were designed to complement sequence between -80 and +25 bases of the initiating AUG codons of the target mRNAs, and the specificities of many were tested by (i) designing different non-overlapping MOs directed against the same mRNA, (ii) injecting MOs differing in five bases, and (iii) performing "rescue" experiments. About 65% of the MOs caused X. tropicalis embryos to develop abnormally (59% of those targeted against novel genes), and we have divided the genes into "synphenotype groups," members of which cause similar loss-of-function phenotypes and that may function in the same developmental pathways. Analysis of the expression patterns of the 202 genes indicates that members of a synphenotype group are not necessarily members of the same synexpression group. This screen provides new insights into early vertebrate development and paves the way for a more comprehensive MO-based analysis of gene function in X. tropicalis.
Rana
Amer Ahmed
AA
Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, United Kingdom.
Collart
Clara
C
Gilchrist
Michael J
MJ
Smith
J C
JC
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D019521:Body Patterning / Q000187:drug effects; D030541:Databases, Genetic; D004625:Embryo, Nonmammalian / Q000002:abnormalities / Q000187:drug effects; D005775:Gastrula / Q000187:drug effects; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D051379:Mice; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*; D010641:Phenotype; D014981:Xenopus / Q000196:embryology* / Q000235:genetics*; D015027:Zebrafish / Q000196:embryology
D016376:Oligonucleotides, Antisense