Figure 2.
Targeted knockdown of gng2 using splicing morpholino in zebrafish embryos. (A) Splice junction morpholino targeted against gng2 exon-intron boundary. (B) RT-PCR of gng2 transcript at tailbud stage in WT and gng2-MO (100 μM) morpholino-injected embryos, comparing cryptic spliced transcript in the morpholino injected embryos to the cDNA and genomic PCR products. (C) Sequencing of the RT-PCR products revealed the misspliced transcript leading to a premature stop (asterisk) and causing a truncation in the protein (exon 1 in bold, exon 2 in plain, and intron in italics and underlined). (D-E) Live morphology of WT control zebrafish embryo (D) and gng2-MO knockdown embryo (E) at 1 dpf. Injection volume was about 1 nL at 1-cell stage embryos. All scale bars are 100 μm.