Interpreting ciliopathy-associated missense variants of uncertain significance (VUS) in an animal model
Interpreting ciliopathy-associated missense variants of uncertain significance (VUS) in an animal model
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解释动物模型中与纤毛病相关的意义不确定的错义变异 (VUS)
DOI：
10.1101/2021.06.17.448799
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Lange K
中科院分区：
文献类型：
--
作者：
Lange K
目的对临床上不能采取有效措施的疾病相关性不确定变异（VUS）的致病性进行研究。在这项研究中，我们探讨了使用一个易处理的动物模型（秀丽隐杆线虫）在体内解释TMEM 67的错义VUS等位基因，纤毛基因与ciliopathies.MethodsCRISPR/Cas9基因编辑用于产生纯合敲入C。携带TMEM 67患者变异体的线虫菌株被工程化到正向同源基因（mks-3）中。如果变异体影响mks-3基因功能，则测量感觉纤毛结构和功能的定量表型测定。通过在人TMEM 67敲除hTERT-RPE 1细胞系中的遗传互补测定来验证蠕虫的结果，所述细胞系测试TMEM 67信号传导功能。elegans准确区分已知的良性（Asp 359 Glu，Thr 360 Ala）和已知的致病性（Glu 361 Ter，Gln 376 Pro）变体。对8个错义VUS的分析表明，3个是良性的（Cys 173 Arg，Thr 176 Ile，Gly 979 Arg），5个是致病性的（Cys 170 Tyr，His 782 Arg，Gly 786 Glu，His 790 Arg，Ser 961 Tyr）。elegans使其成为易于处理的体内动物模型，其允许快速、成本有效地解释纤毛病相关的错义VUS等位基因。
PurposeBetter methods are required to interpret the pathogenicity of disease-associated variants of uncertain significance (VUS), which cannot be actioned clinically. In this study, we explore the use of a tractable animal model (Caenorhabditis elegans) for in vivo interpretation of missense VUS alleles of TMEM67, a cilia gene associated with ciliopathies.MethodsCRISPR/Cas9 gene editing was used to generate homozygous knock-in C. elegans worm strains carrying TMEM67 patient variants engineered into the orthologous gene (mks-3). Quantitative phenotypic assays of sensory cilia structure and function measured if the variants affect mks-3 gene function. Results from worms were validated by a genetic complementation assay in a human TMEM67 knock-out hTERT-RPE1 cell line that tests a TMEM67 signaling function.ResultsAssays in C. elegans accurately distinguished between known benign (Asp359Glu, Thr360Ala) and known pathogenic (Glu361Ter, Gln376Pro) variants. Analysis of eight missense VUS generated evidence that three are benign (Cys173Arg, Thr176Ile, Gly979Arg) and five are pathogenic (Cys170Tyr, His782Arg, Gly786Glu, His790Arg, Ser961Tyr).ConclusionEfficient genome editing and quantitative functional assays in C. elegans make it a tractable in vivo animal model that allows rapid, cost-effective interpretation of ciliopathy-associated missense VUS alleles.
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CiliaCarta：完整且经过验证的纤毛基因简编
影响因子：
3.7
作者：
Teunis J. P. van Dam;Julie E. Kennedy;Robin van der Lee;E. Vrieze;K. Wunderlich;S. Rix;G. Dougherty;N. Lambacher;Chunmei Li;Victor L Jensen;M. Leroux;R. Hjeij;N. Horn;Y. Texier;Yasmin Wissinger;J. van Reeuwijk;G. Wheway;B. Knapp;J. Scheel;B. Franco;D. Mans;E. van Wijk;F. Képès;G. Slaats;G. Toedt;H. Kremer;H. Omran;K. Szymanska;K. Koutroumpas;M. Ueffing;Thanh;S. Letteboer;M. Oud;S. V. van Beersum;M. Schmidts;P. Beales;Q. Lu;R. Giles;R. Szklarczyk;R. Russell;T. Gibson;Colin A. Johnson;O. Blacque;U. Wolfrum;K. Boldt;R. Roepman;Victor Hernandez;M. Huynen
通讯作者：
M. Huynen
影响因子：
3.9
作者：
Tallila, Jonna;Salonen, Riitta;Kohlschmidt, Nicolai;Peltonen, Leena;Kestila, Marjo
通讯作者：
Kestila, Marjo
影响因子：
5.6
作者：
Zhu B;Mak JCH;Morris AP;Marson AG;Barclay JW;Sills GJ;Morgan A
通讯作者：
Morgan A
影响因子：
3.3
作者：
T. Starich;R. K. Herman;C. K. Kari;Wen-Hui Yeh;Wendy S. Schackwitz;Marcia W. Schuyler;J. Collet
通讯作者：
T. Starich;R. K. Herman;C. K. Kari;Wen-Hui Yeh;Wendy S. Schackwitz;Marcia W. Schuyler;J. Collet
DOI：
10.1007/978-1-4939-2842-2_14
发表时间：
2015-01-01
期刊：
C. ELEGANS: METHODS AND APPLICATIONS, 2ND EDITION
影响因子：
--
作者：
Churgin, Matthew A.;Fang-Yen, Christopher
通讯作者：
Fang-Yen, Christopher