Reverse genetics in zebrafish.
The zebrafish has become a popular model system for the study of vertebrate developmental biology because of its numerous strengths as a molecular genetic and embryological system. To determine the requirement for specific genes during embryogenesis, it is necessary to generate organisms carrying loss-of-function mutations. This can be accomplished in zebrafish through a reverse genetic approach. This review discusses the current techniques for generating mutations in known genes in zebrafish. These techniques include the generation of chromosomal deletions and the subsequent identification of complementation groups within deletions through noncomplementation assays. In addition, this review will discuss methods currently being evaluated that may improve the methods for finding mutations in a known sequence, including screening for randomly induced small deletions within genes and screening for randomly induced point mutations within specific genes.
Lekven
A C
AC
Howard Hughes Medical Institute, Department of Pharmacology, Center for Developmental Biology, University of Washington School of Medicine, Seattle, Washington 98195, USA. alekven@u.washington.edu
Helde
K A
KA
Thorpe
C J
CJ
Rooke
R
R
Moon
R T
RT
D000818:Animals; D001755:Blastocyst / Q000528:radiation effects; D002872:Chromosome Deletion; D004252:DNA Mutational Analysis / Q000379:methods; D018390:Gene Targeting; D005816:Genetic Complementation Test; D005821:Genetic Techniques; D005820:Genetic Testing / Q000379:methods; D020180:Heteroduplex Analysis; D008954:Models, Biological; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction; D017384:Sequence Deletion; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*