Capped mRNAs were synthesized in vitro for pinhead, pinhead-HA, pinhead-GFP, mCherry-CAAX, admp, bmp2b, and gfp from the corresponding linearized plasmids using the mMessage mMachine kit (Ambion). The following MOs were synthesized by Gene Tools and resuspended in nuclease-free water: ph MO (5′-ACTGACAGCAGTAAATCCATAGTCC-3′), admp MO (ad MO; 5′-TGGACAACATTGTAAAGAACATTGC-3′) (14), bmp2b MO (5′-CGCGGACCACGGCGACCATGATC-3′) (30), and chd MO (5′-ATCCACAGCAGCCCCTCCATCATCC-3′) (29). The mRNA and MOs were injected into the yolk, while plasmid DNA was injected into the cytoplasm of one-cell stage embryos.

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