METHODS: Sanger sequencing was used for pedigree verification, and the effects of this variant on mRNA splicing were analyzed through minigene assay.
RESULTS: The study revealed that this variant led to the creation of a previously unreported splice site in the fourth intron, resulting in the incorporation of an 11 bp sequence from the intron into the final transcript. This alteration led to a frameshift and formation of a premature termination codon, impacting the structure of the aggrecan protein.
CONCLUSIONS: We document the pathogenicity of an ACAN non-canonical splicing-site variant, emphasizing the significance of considering intronic variants during genetic testing.
方法:使用Sanger测序进行系谱验证,并通过小基因分析分析了该变体对mRNA剪接的影响。
结果:研究表明,该变体导致在第四个内含子中产生了以前未报道的剪接位点,导致来自内含子的11bp序列掺入到最终的转录物中。这种改变导致移码和过早终止密码子的形成,影响聚集蛋白聚糖蛋白的结构。
结论:我们记录了ACAN非规范剪接位点变体的致病性,强调在基因测试中考虑内含子变异的重要性。