Current Protocols in Human Genetics最新文献_第9页

Detecting APC Gene Mutations in Familial Adenomatous Polyposis (FAP) 家族性腺瘤性息肉病(FAP) APC基因突变检测

Current Protocols in Human Genetics Pub Date : 2017-01-11 DOI: 10.1002/cphg.29

Babi Ramesh Reddy Nallamilli, Madhuri Hegde

{"title":"Detecting APC Gene Mutations in Familial Adenomatous Polyposis (FAP)","authors":"Babi Ramesh Reddy Nallamilli, Madhuri Hegde","doi":"10.1002/cphg.29","DOIUrl":"10.1002/cphg.29","url":null,"abstract":"Hereditary forms of colorectal cancer (CRC) account for up to 5% of total cases. Familial adenomatous polyposis (FAP) is an autosomal dominant condition affecting nearly 1 in 5000 people and accounts for only about 1% of all CRCs. It is characterized by the progressive development of hundreds to thousands of adenomatous colon polyps. The gene associated with FAP (APC) contains 15 coding exons. The mutation spectrum of the APC gene is broad in that 87% of causative mutations are point mutations (including other sequence variants) and around 10% to 15% are intragenic deletions and duplications. The strategy for molecular diagnostic testing for FAP involves initial full sequence analysis of APC for sequence variants followed by screening for deletion/duplications using microarray-based comparative genomic hybridization (array CGH) or Multiplex Ligation-dependent Probe Amplification (MLPA). Recently, next generation sequencing (NGS)-based targeted gene analysis has become clinically available for detection of point mutations and other sequence variants. This unit discusses detailed protocols for an NGS-based sequencing assay, PCR-based Sanger sequencing, and array CGH. © 2017 by John Wiley & Sons, Inc.","PeriodicalId":40007,"journal":{"name":"Current Protocols in Human Genetics","volume":"92 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2017-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cphg.29","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83965045","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 18

Generating Exome Enriched Sequencing Libraries from Formalin-Fixed, Paraffin-Embedded Tissue DNA for Next-Generation Sequencing 从福尔马林固定石蜡包埋的组织DNA中生成外显子组富集测序文库，用于下一代测序

Current Protocols in Human Genetics Pub Date : 2017-01-11 DOI: 10.1002/cphg.27

Beth A. Marosy, Brian D. Craig, Kurt N. Hetrick, P. Dane Witmer, Hua Ling, Sean M. Griffith, Benjamin Myers, Elaine A. Ostrander, Janet L. Stanford, Lawrence C. Brody, Kimberly F. Doheny

引用次数: 5

Highly Expandable Human iPS Cell–Derived Neural Progenitor Cells (NPC) and Neurons for Central Nervous System Disease Modeling and High-Throughput Screening 用于中枢神经系统疾病建模和高通量筛选的高度可扩展的人类iPS细胞衍生神经祖细胞(NPC)和神经元

Current Protocols in Human Genetics Pub Date : 2017-01-11 DOI: 10.1002/cphg.33

Chialin Cheng, Daniel M. Fass, Kat Folz-Donahue, Marcy E. MacDonald, Stephen J. Haggarty

{"title":"Highly Expandable Human iPS Cell–Derived Neural Progenitor Cells (NPC) and Neurons for Central Nervous System Disease Modeling and High-Throughput Screening","authors":"Chialin Cheng, Daniel M. Fass, Kat Folz-Donahue, Marcy E. MacDonald, Stephen J. Haggarty","doi":"10.1002/cphg.33","DOIUrl":"10.1002/cphg.33","url":null,"abstract":"Reprogramming of human somatic cells into induced pluripotent stem (iPS) cells has greatly expanded the set of research tools available to investigate the molecular and cellular mechanisms underlying central nervous system (CNS) disorders. Realizing the promise of iPS cell technology for the identification of novel therapeutic targets and for high-throughput drug screening requires implementation of methods for the large-scale production of defined CNS cell types. Here we describe a protocol for generating stable, highly expandable, iPS cell–derived CNS neural progenitor cells (NPC) using multi-dimensional fluorescence activated cell sorting (FACS) to purify NPC defined by cell surface markers. In addition, we describe a rapid, efficient, and reproducible method for generating excitatory cortical-like neurons from these NPC through inducible expression of the pro-neural transcription factor Neurogenin 2 (iNgn2-NPC). Finally, we describe methodology for the use of iNgn2-NPC for probing human neuroplasticity and mechanisms underlying CNS disorders using high-content, single-cell-level automated microscopy assays. © 2017 by John Wiley & Sons, Inc.","PeriodicalId":40007,"journal":{"name":"Current Protocols in Human Genetics","volume":"92 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2017-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cphg.33","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73498680","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 34

Molecular Analysis of Gene Rearrangements and Mutations in Acute Leukemias and Myeloid Neoplasms 急性白血病和髓系肿瘤基因重排和突变的分子分析

Current Protocols in Human Genetics Pub Date : 2017-01-11 DOI: 10.1002/cphg.31

Lynette M. Sholl, Janina Longtine, Frank C. Kuo

引用次数: 4

Human Induced Pluripotent Stem (hiPS) Cells from Urine Samples: A Non-Integrative and Feeder-Free Reprogramming Strategy 来自尿液样本的人类诱导多能干细胞(hiPS):一种非整合和无喂食的重编程策略

Current Protocols in Human Genetics Pub Date : 2017-01-11 DOI: 10.1002/cphg.26

Clara Steichen, Karim Si-Tayeb, Fanny Wulkan, Thayane Crestani, Graça Rosas, Rafael Dariolli, Alexandre C. Pereira, Jose E. Krieger

{"title":"Human Induced Pluripotent Stem (hiPS) Cells from Urine Samples: A Non-Integrative and Feeder-Free Reprogramming Strategy","authors":"Clara Steichen, Karim Si-Tayeb, Fanny Wulkan, Thayane Crestani, Graça Rosas, Rafael Dariolli, Alexandre C. Pereira, Jose E. Krieger","doi":"10.1002/cphg.26","DOIUrl":"10.1002/cphg.26","url":null,"abstract":"Human induced pluripotent stem (hiPS) cell technology has already revolutionized some aspects of fundamental and applied research such as study of disease mechanisms and pharmacology screening. The first clinical trial using hiPS cell-derived cells began in Japan, only 10 years after the publication of the proof-of concept article. In this exciting context, strategies to generate hiPS cells have evolved quickly, tending towards non-invasive protocols to sample somatic cells combined with “safer” reprogramming strategies. In this unit, we describe a protocol combining both of these advantages to generate hiPS cells with episomal plasmid transfection from urine samples of individuals carrying the desired genotype. Based on previous published works, this simplified protocol requires minimal equipment and reagents, and is suitable both for scientists familiar with the hiPS cells technology and neophytes. HiPS cells displaying classical features of pluripotency and suitable for all desired downstream applications are generated rapidly (<10 weeks) and with high efficiency. © 2017 by John Wiley & Sons, Inc.","PeriodicalId":40007,"journal":{"name":"Current Protocols in Human Genetics","volume":"92 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2017-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cphg.26","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89158872","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 16

Culturing and Neuronal Differentiation of Human Dental Pulp Stem Cells 人牙髓干细胞的培养与神经元分化

Current Protocols in Human Genetics Pub Date : 2017-01-11 DOI: 10.1002/cphg.28

Sarita Goorha, Lawrence T. Reiter

引用次数: 31

Diagnosis of Spinocerebellar Ataxias Caused by Trinucleotide Repeat Expansions 三核苷酸重复扩增引起脊髓小脑共济失调的诊断

Current Protocols in Human Genetics Pub Date : 2017-01-11 DOI: 10.1002/cphg.30

Joanne E. Martindale

引用次数: 3

Assay for Transposase-Accessible Chromatin Using Sequencing (ATAC-seq) Data Analysis 使用测序(ATAC-seq)数据分析转座酶可及染色质的测定

Current Protocols in Human Genetics Pub Date : 2017-01-11 DOI: 10.1002/cphg.32

Kristy L.S. Miskimen, E. Ricky Chan, Jonathan L. Haines

引用次数: 11

Fabry Disease Biomarkers: Analysis of Urinary Lyso-Gb3 and Seven Related Analogs Using Tandem Mass Spectrometry 法布里病生物标志物:用串联质谱法分析尿溶索- gb3和7种相关类似物

Current Protocols in Human Genetics Pub Date : 2016-07-01 DOI: 10.1002/cphg.1

Pamela Lavoie, Michel Boutin, Mona Abaoui, Christiane Auray-Blais

{"title":"Fabry Disease Biomarkers: Analysis of Urinary Lyso-Gb3 and Seven Related Analogs Using Tandem Mass Spectrometry","authors":"Pamela Lavoie, Michel Boutin, Mona Abaoui, Christiane Auray-Blais","doi":"10.1002/cphg.1","DOIUrl":"10.1002/cphg.1","url":null,"abstract":"Fabry disease is an X-linked lysosomal storage disorder caused by the absence or reduction of the enzyme α-galactosidase A activity. Currently, globotriaosylsphingosine (lyso-Gb3) and globotriaosylceramide (Gb3) are used as biomarkers to diagnose and monitor Fabry patients. However, recent metabolomic studies have shown that several glycosphingolipids are also elevated in biological fluids of affected patients and may be related to disease manifestations. This unit describes a multiplex methodology targeting the analysis of urinary lyso-Gb3 and seven structurally related analogs. A solid-phase extraction process is performed, then lyso-Gb3 and its analogs are analyzed simultaneously with an internal standard by ultra-performance liquid chromatography (UPLC) coupled to a tandem mass spectrometry (MS/MS) system. This methodology can be useful for the diagnosis of Fabry patients, including patients with cardiac variant mutations, but also to monitor the efficacy of therapeutic interventions, considering that lyso-Gb3 analogs are more elevated than lyso-Gb3 itself in urine. © 2016 by John Wiley & Sons, Inc.","PeriodicalId":40007,"journal":{"name":"Current Protocols in Human Genetics","volume":"90 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cphg.1","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34693733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 9

Sequence Variant Descriptions: HGVS Nomenclature and Mutalyzer 序列变异描述:HGVS命名法和变异分析仪

Current Protocols in Human Genetics Pub Date : 2016-07-01 DOI: 10.1002/cphg.2

Johan T. den Dunnen

引用次数: 42