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Fig. 3 Knockdown of abca12 and snap29 expression by morpholinos. (A) Knockdown of abca12. (B) Knockdown of snap29. MO1 and MO2 morpholinos (right lanes, the upper panel), which target the splice donor site at the exon-4–intron-4 border of the corresponding genes, prevents pre-mRNA splicing. The consequences of MO1 on abca12 pre-mRNA splicing and MO2 on snap29 mRNA splicing were determined by RT-PCR. The results showed the retention of intron 4 in the majority of mRNA transcripts (>90%) as compared with the normally transcribed control. The mRNA levels were normalized by the level of β-actin mRNA (lower panels). cDNA and gDNA represent amplification of the corresponding complementary DNA and genomic DNA, respectively. Injections with the 5-bp mismatched control morpholino for abca12 (cMO) or global standard control morpholino (scMO) did not alter pre-mRNA processing, similar to the uninjected controls or those injected with phenol red.