Molecular diagnosis of citrin deficiency in an infant with intrahepatic cholestasis: identification of a 21.7kb gross deletion that completely silences the transcriptional and translational expression of the affected SLC25A13 allele.
Molecular diagnosis of citrin deficiency in an infant with intrahepatic cholestasis: identification of a 21.7kb gross deletion that completely silences the transcriptional and translational expression of the affected SLC25A13 allele.
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肝内胆汁淤积婴儿柑橘缺乏症的分子诊断：鉴定出 21.7kb 的总缺失，该缺失完全沉默了受影响的 SLC25A13 等位基因的转录和翻译表达
DOI：
10.18632/oncotarget.19901
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发表时间：
2017-10-20
期刊：
影响因子：
--
通讯作者：
Song YZ
中科院分区：
文献类型：
--
作者：
Zhang ZH;Lin WX;Zheng QQ;Guo L;Song YZ
新生儿胆汁淤积症(NICCD)是由双等位基因SLC25A13突变引起的，SLC25A13分析为NICCD的确诊提供了可靠的依据。然而，仅通过传统的DNA分析不能检测到该基因中的新的大插入/缺失。这项研究旨在探索高度怀疑患有NICCD的婴儿的明确诊断证据。SLC25A13基因的流行突变筛选和Sanger测序发现了一个父系遗传突变C.851_854del4。然而，在Western blotting和cDNA克隆分析中，分别没有检测到Citrin蛋白和SLC25A13的母源转录本。在此基础上，利用SNP分析和半定量聚合酶链式反应对隐藏的母体突变进行了精确定位，最终确定为一个新的大缺失c.-3251_c.15+18443del21709bp，涉及SLC25A13启动子区域和翻译起始密码子所在的整个外显子1。因此，NICCD肯定是在婴儿身上诊断出来的。据我们所知，SLC25A13等位基因转录和翻译表达的沉默，是迄今为止SLC25A13突变谱中最大的缺失。特别值得注意的是，使用从扩增的外周血淋巴细胞中提取的线粒体蛋白的Western blotting方法，可能是诊断NICCD的一种新的微创和更可行的分子工具。
Neonatal Intrahepatic Cholestasis caused by Citrin Deficiency (NICCD) arises from biallelic SLC25A13 mutations, and SLC25A13 analysis provides reliable evidences for NICCD definite diagnosis. However, novel large insertions/deletions in this gene could not be detected just by conventional DNA analysis. This study aimed to explore definite diagnostic evidences for an infant highly-suspected to have NICCD. Prevalent mutation screening and Sanger sequencing of SLC25A13 gene just revealed a paternally-inherited mutation c.851_854del4. Nevertheless, neither citrin protein nor SLC25A13 transcripts of maternal origin could be detected on Western blotting and cDNA cloning analysis, respectively. On this basis, the hidden maternal mutation was precisely positioned using SNP analysis and semi-quantitative PCR, and finally identified as a novel large deletion c.-3251_c.15+18443del21709bp, which involved the SLC25A13 promoter region and the entire exon 1 where locates the translation initiation codon. Hence, NICCD was definitely diagnosed in the infant. To the best of our knowledge, the novel gross deletion, which silenced the transcriptional and translational expression of the affected SLC25A13 allele, is the hitherto largest deletion in SLC25A13 mutation spectrum. The Western blotting approach using mitochondrial protein extracted from expanded peripheral blood lymphocytes, of particular note, might be a new minimally-invasive and more-feasible molecular tool for NICCD diagnosis.
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中国柑橘缺乏症儿科患者的分子诊断：SLC25A13 突变谱和地理分布
影响因子：
4.6
作者：
Lin WX;Zeng HS;Zhang ZH;Mao M;Zheng QQ;Zhao ST;Cheng Y;Chen FP;Wen WR;Song YZ
通讯作者：
Song YZ
影响因子：
3.7
作者：
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通讯作者：
Williams, Kenneth C.
影响因子：
8
作者：
Dimmock, David;Kobayashi, Keiko;Scaglia, Fernando
通讯作者：
Scaglia, Fernando
影响因子：
4.2
作者：
Hutchin, T.;Preece, M. A.;Baumann, U.
通讯作者：
Baumann, U.
DOI：
10.1016/j.bbamcr.2016.04.011
发表时间：
2016-10-01
影响因子：
5.1
作者：
Amoedo, N. D.;Punzi, G.;Rossignol, R.
通讯作者：
Rossignol, R.