Insertional mutagenesis in zebrafish rapidly identifies genes essential for early vertebrate development.
To rapidly identify genes required for early vertebrate development, we are carrying out a large-scale, insertional mutagenesis screen in zebrafish, using mouse retroviral vectors as the mutagen. We will obtain mutations in 450 to 500 different genes--roughly 20% of the genes that can be mutated to produce a visible embryonic phenotype in this species--and will clone the majority of the mutated alleles. So far, we have isolated more than 500 insertional mutants. Here we describe the first 75 insertional mutants for which the disrupted genes have been identified. In agreement with chemical mutagenesis screens, approximately one-third of the mutants have developmental defects that affect primarily one or a small number of organs, body shape or swimming behavior; the rest of the mutants show more widespread or pleiotropic abnormalities. Many of the genes we identified have not been previously assigned a biological role in vivo. Roughly 20% of the mutants result from lesions in genes for which the biochemical and cellular function of the proteins they encode cannot be deduced with confidence, if at all, from their predicted amino-acid sequences. All of the genes have either orthologs or clearly related genes in human. These results provide an unbiased view of the genetic construction kit for a vertebrate embryo, reveal the diversity of genes required for vertebrate development and suggest that hundreds of genes of unknown biochemical function essential for vertebrate development have yet to be identified.
Golling
Gregory
G
Center for Cancer Research and Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
Amsterdam
Adam
A
Sun
Zhaoxia
Z
Antonelli
Marcelo
M
Maldonado
Ernesto
E
Chen
Wenbiao
W
Burgess
Shawn
S
Haldi
Maryann
M
Artzt
Karen
K
Farrington
Sarah
S
Lin
Shuh-Yow
SY
Nissen
Robert M
RM
Hopkins
Nancy
N
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D004625:Embryo, Nonmammalian / Q000196:embryology / Q000502:physiology; D018507:Gene Expression Regulation, Developmental; D016254:Mutagenesis, Insertional / Q000235:genetics*; D009154:Mutation; D012190:Retroviridae / Q000235:genetics; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*