Production of maternal-zygotic mutant zebrafish by germ-line replacement.
We report a generally applicable strategy for transferring zygotic lethal mutations through the zebrafish germ line. By using a morpholino oligonucleotide that blocks primordial germ cell (PGC) development, we generate embryos devoid of endogenous PGCs to serve as hosts for the transplantation of germ cells derived from homozygous mutant donors. Successful transfers are identified by the localization of specifically labeled donor PGCs to the region of the developing gonad in chimeric embryos. This strategy, which results in the complete replacement of the host germ line with donor PGCs, was validated by the generation of maternal and maternal-zygotic mutants for the miles apart locus. This germ-line replacement technique provides a powerful tool for studying the maternal effects of zygotic lethal mutations. Furthermore, the ability to generate large clutches of purely mutant embryos will greatly facilitate embryological, genetic, genomic, and biochemical studies.
Ciruna
Brian
B
Developmental Genetics Program, Skirball Institute of Biomolecular Medicine, and Department of Cell Biology, New York University School of Medicine, New York 10016, USA.
Weidinger
Gilbert
G
Knaut
Holger
H
Thisse
Bernard
B
Thisse
Christine
C
Raz
Erez
E
Schier
Alexander F
AF
D000818:Animals; D001483:Base Sequence; D017690:Cell Transplantation; D017931:DNA Primers; D005260:Female; D018392:Genomic Imprinting; D005838:Genotype; D018095:Germ-Line Mutation; D008297:Male; D012333:RNA, Messenger / Q000235:genetics; D018183:Transplantation Chimera; D015027:Zebrafish / Q000235:genetics*; D015053:Zygote / Q000502:physiology*
D017931:DNA Primers; D012333:RNA, Messenger