Xenbase Image ID: 134571

Figure 4. Morpholinos (MOs) directed against 25H12.1 (pinhead) splice sites cause aberrant splicing and an increase in detectable pinhead mRNA. A: Diagram of part of the 25H12.1 gene in X. tropicalis with exonic sequences represented as boxes and intronic sections as black lines. I: The target sites of 25H12.1moA and 25H12.1moB at the boundary of a 179-bp intron and a 96-bp exon are shown as moA and moB. Primers f2 and r1 used for reverse transcriptase-polymerase chain reaction (RT-PCR) are depicted as arrows. II: Wild-type cDNA yields a 187-bp RT-PCR product. III,IV: Aberrantly spliced cDNAs yield 366-bp and 91-bp RT-PCR products with f2 and r1 due to inclusion of the 179-bp intron or skipping of the 96-bp exon. The gel shows RT-PCR (primers f2 and r1) of 25H12.1 cDNA from uninjected (WT, wild-type) neurula embryos or embryos injected at one-cell stage with 10 ng of 25H12.moA (moA) or 25H12.1moB (moB). Amplified DNA fragments are labelled (I–IV) according to the diagram shown above. Both MOs induce abnormal splicing. M, marker lane; C, no cDNA control; G, Xenopus tropicalis genomic DNA; P, plasmid DNA from TGas046h20 that includes insertion of the 179-bp intron. B:X. tropicalis embryos uninjected (a) or injected at the one-cell stage with 10 ng of 25H12.1moB (b,c) and hybridised with an 25H12.1 antisense RNA probe. As well as the pinhead phenotype, injected embryos show increased amounts of 25H12.1 mRNA in anterior regions (arrowheads in b) and around the tail bud (arrowhead in c). Image published in: Kenwrick S et al. (2004) Copyright © 2004. Image reproduced with permission of the Publisher, John Wiley & Sons. Gene Synonyms Species Stage(s) Tissue pnhd pinhead X. tropicalis Throughout NF stage 24   Image source: Published Experiment + Assay Source Phenotypes and Disease Xtr Wt + pnhd MO + NF24 (morphology) fig.4.B.b,c Anatomical Phenotype abnormally posterioralized embryo decreased width of the head Larger Image Printer Friendly View

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