Germ-line replacement strategy using a GFP-nos1-3′UTR labeling technique. (A) Overview of transplantation strategy showing the transfer of cells from the margin of GFP-nos1-3′UTR-labeled mutant donor embryos into the margin of αPGC-MO-injected WT hosts. "m" refers to any zygotic mutation, in this case mil^te273. (B) At 30 hpf, PGCs from injected control mil_ix embryos are specifically labeled with GFP (arrowhead). (C) Host embryos were screened at 30 hpf for the presence of GFP-labeled donor PGCs and their normal migration into the gonadal mesoderm at the anterior region of the yolk extension (arrow).

Germ-line replacement strategy using the rhodamine-dextran labeling technique. (A) Overview of transplantation strategy showing the transfer of cells from the margin of rhodamine-dextran-labeled mutant donor embryos into the animal pole of αPGC-MO-injected WT hosts. (B) At 30 hpf, all cells within control donor embryos are efficiently labeled with rhodamine-dextran. (C) Chimeric host embryo at 30 hpf showing somatic contribution of rhodamine-dextran-labeled donor cells to anterior neuroectoderm lineages (*). Host embryos were screened for the presence of labeled donor PGCs that had migrated successfully into the gonadal mesoderm (arrow).

PCR genotyping of mil-mutant and WT embryos. Shown is 3% agarose gel resolution of HpyCh4V and AciI digestion products of PCR fragments amplified from genomic DNA prepared from mil^te273/mil^te273, mil^te273/+, and +/+ embryos.