A Mismatch EndoNuclease Array-Based Methodology (MENA) for Identifying Known SNPs or Novel Point Mutations.
A Mismatch EndoNuclease Array-Based Methodology (MENA) for Identifying Known SNPs or Novel Point Mutations.
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DOI：
10.3390/microarrays5020007
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发表时间：
2016-04-05
期刊：
影响因子：
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通讯作者：
Manak, J Robert
中科院分区：
文献类型：
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作者：
Comeron, Josep M;Reed, Jordan;Manak, J Robert
准确快速地识别或确认单核苷酸多态性（SNP）、点突变和其他人类基因组变异有助于了解疾病的遗传基础。我们开发了一种新方法（称为 MENA（错配核酸内切酶阵列）），将 DNA 错配核酸内切酶学与平铺微阵列杂交配对，以便对已知点突变（例如 SNP）进行基因分型，并识别以前未发现的点突变和小插入缺失。我们表明，除了以低至 10% 的错误发现率识别新的点突变和小插入缺失之外，我们的检测还可以快速对人类基因组 DNA 样本中已知的 SNP 进行基因分型，准确度高达 99%。我们的技术为各种应用提供了一个平台，包括：（1）对已知的 SNP 进行基因分型以及通过全基因组测序分析确认新发现的 SNP； (2) 在可获得基因组序列信息的任何生物体的任何基因组区域中识别新的点突变和插入缺失； (3) 筛选与患者样本中特定疾病和病症相关的基因组，以识别致病突变。作为使用 MENA 发现新突变的原理证明，我们报告了果蝇中贝多芬 (btv) 基因的新等位基因的鉴定，该等位基因编码对听觉机械感觉重要的纤毛细胞质动力蛋白运动蛋白。
Accurate and rapid identification or confirmation of single nucleotide polymorphisms (SNPs), point mutations and other human genomic variation facilitates understanding the genetic basis of disease. We have developed a new methodology (called MENA (Mismatch EndoNuclease Array)) pairing DNA mismatch endonuclease enzymology with tiling microarray hybridization in order to genotype both known point mutations (such as SNPs) as well as identify previously undiscovered point mutations and small indels. We show that our assay can rapidly genotype known SNPs in a human genomic DNA sample with 99% accuracy, in addition to identifying novel point mutations and small indels with a false discovery rate as low as 10%. Our technology provides a platform for a variety of applications, including: (1) genotyping known SNPs as well as confirming newly discovered SNPs from whole genome sequencing analyses; (2) identifying novel point mutations and indels in any genomic region from any organism for which genome sequence information is available; and (3) screening panels of genes associated with particular diseases and disorders in patient samples to identify causative mutations. As a proof of principle for using MENA to discover novel mutations, we report identification of a novel allele of the beethoven (btv) gene in Drosophila, which encodes a ciliary cytoplasmic dynein motor protein important for auditory mechanosensation.