Current Protocols in Human Genetics最新文献_第4页

Glucose-Responsiveness of Pancreatic β-Like (GRP β-L) Cells Generated from Human Pluripotent Stem Cells 人多能干细胞生成胰腺β样细胞(GRP β-L)的葡萄糖反应性

Current Protocols in Human Genetics Pub Date : 2018-10-18 DOI: 10.1002/cphg.71

Bahareh Rajaei, Mohammad Massumi, Michael Wheeler

{"title":"Glucose-Responsiveness of Pancreatic β-Like (GRP β-L) Cells Generated from Human Pluripotent Stem Cells","authors":"Bahareh Rajaei, Mohammad Massumi, Michael Wheeler","doi":"10.1002/cphg.71","DOIUrl":"10.1002/cphg.71","url":null,"abstract":"The International Diabetic Federation estimated that 415 million adults currently have diabetes and 318 million adults had impaired glucose tolerance, putting them at high risk of developing diabetes in the future. In Type 1 Diabetes (T1D), the β cells are lost because of autoimmune reactions. Although islet transplantation has been a promising therapy for T1D, it is greatly limited by pancreatic donors. Here, we describe a protocol to generate glucose- responsive pancreatic β-like (GRPβ-L) cells from human-induced pluripotent stem (iPS) cells. We recapitulate in vivo pancreas development by in vitro induction of differentiating human (iPS) cells with stage-specific signaling molecules and proteins. Inhibition of Tyrosine Kinase receptor AXL, TGF-β, and Notch signaling pathways in the final stage of the five-stage protocol could efficiently generate GRPβ-L from the endocrine progenitor. Differentiation of human iPS cells through the protocol could result in functional GRPβ-L cells, which could be used in pharmaceutical and β cell biology studies. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":40007,"journal":{"name":"Current Protocols in Human Genetics","volume":"100 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cphg.71","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10379419","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: 2

A Guided Protocol for Array Based T2C: A High-Quality Selective High-Resolution High-Throughput Chromosome Interaction Capture. 基于阵列的T2C指导协议:高质量选择性高分辨率高通量染色体相互作用捕获。

Current Protocols in Human Genetics Pub Date : 2018-10-01 Epub Date: 2018-09-10 DOI: 10.1002/cphg.55

Tobias A Knoch

{"title":"A Guided Protocol for Array Based T2C: A High-Quality Selective High-Resolution High-Throughput Chromosome Interaction Capture.","authors":"Tobias A Knoch","doi":"10.1002/cphg.55","DOIUrl":"https://doi.org/10.1002/cphg.55","url":null,"abstract":"After now more than 170 years of research the dynamic three-dimensional chromatin architecture of genomes and the co-evolved interaction networks of regulatory elements which create genome function - i.e. the storage, expression, and finally replication of genetic information - involves ever more investigative efforts in respect to not only the pure understanding of living organisms, but also diagnosis, treatment, and even future genome engineering. To study genomic interactions, we developed a novel and superior high-quality selective high-resolution, high-throughput chromosome interaction capture method - T2C (targeted chromatin capture) - which allows to arbitrarily balance resolution, frequency range of interactions, and the investigated general genetic region or single interactions in a highly cost-effective manner in respect to the obtainable result and compared to other techniques. Beyond, T2C has such a high signal-to-noise ratio at high resolution that the \"genomic\" statistical mechanics level can be reached. With the guided T2C protocol described here, we were already able to finally determine the chromatin quasi-fiber conformation and its folding into stable multi-loop aggregates/rosettes connected by a linker. Actually, this guided T2C protocol provides the means for architectural genome sequencing from the level of the single base pair to the entire cell nucleus and thus to analyze genetic interactions in respect to genome function in a systems biological manner in general as well as in settings ranging from basic research, via diagnostics and treatment, to genome engineering. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":40007,"journal":{"name":"Current Protocols in Human Genetics","volume":" ","pages":"e55"},"PeriodicalIF":0.0,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cphg.55","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36477300","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: 2

Multicolor Fluorescence In Situ Hybridization (FISH) Approaches for Simultaneous Analysis of the Entire Human Genome. 同时分析整个人类基因组的多色荧光原位杂交(FISH)方法。

Current Protocols in Human Genetics Pub Date : 2018-10-01 Epub Date: 2018-09-14 DOI: 10.1002/cphg.70

Chengsheng Zhang, Eliza Cerveira, Willem Rens, Fengtang Yang, Charles Lee

引用次数: 5

MicroRNA Isolation from Plasma for Real-Time qPCR Array. 实时荧光定量pcr阵列从血浆中分离MicroRNA。

Current Protocols in Human Genetics Pub Date : 2018-10-01 Epub Date: 2018-09-14 DOI: 10.1002/cphg.69

Isabel Witvrouwen, Andreas B Gevaert, Emeline M Van Craenenbroeck, Amaryllis H Van Craenenbroeck

引用次数: 3

Issue Information TOC 发布信息TOC

Current Protocols in Human Genetics Pub Date : 2018-10-01 DOI: 10.1002/cphg.73

引用次数: 0

Pancreatic Beta Cell Differentiation From Human Pluripotent Stem Cells. 胰岛β细胞从人多能干细胞分化。

Current Protocols in Human Genetics Pub Date : 2018-10-01 DOI: 10.1002/cphg.68

Lina Sui, Rudolph L Leibel, Dieter Egli

{"title":"Pancreatic Beta Cell Differentiation From Human Pluripotent Stem Cells.","authors":"Lina Sui, Rudolph L Leibel, Dieter Egli","doi":"10.1002/cphg.68","DOIUrl":"https://doi.org/10.1002/cphg.68","url":null,"abstract":"Insulin-expressing beta cells are crucial for the maintenance of systemic glucose homeostasis. Elucidation of the molecular and cellular mechanisms of beta cell development, expansion, survival, and function are required for full understanding of the molecular pathogenesis of diabetes. However, access to human beta cells for such studies is limited by virtue of the logistics of acquisition, prior medical status of donor, and imperfect culture systems for maintaining beta cell identity and function after isolation from human pancreas. Here, a technique for generation of beta cells from human pluripotent stem cells (hPSCs) by modification of key signaling pathways during islet development is described. Up to 70% C-peptide-positive beta cells can be obtained from endodermal anlagen after 27 days of differentiation with specific growth factors and small molecules. Although 50% of them are monohormonal C-peptide-positive cells and have molecular and cellular characteristics consistent with human beta cells in the Islets of Langerhans, a sub-population co-expressing other endocrine markers are also generated, indicating the immaturity of these cells. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":40007,"journal":{"name":"Current Protocols in Human Genetics","volume":"99 1","pages":"e68"},"PeriodicalIF":0.0,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cphg.68","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10752705","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: 17

Quantification of Muscle Contraction In Vitro and In Vivo Using MUSCLEMOTION Software: From Stem Cell-Derived Cardiomyocytes to Zebrafish and Human Hearts. 使用MUSCLEMOTION软件量化体外和体内肌肉收缩:从干细胞衍生的心肌细胞到斑马鱼和人类心脏。

Current Protocols in Human Genetics Pub Date : 2018-10-01 Epub Date: 2018-09-25 DOI: 10.1002/cphg.67

Berend J van Meer, Luca Sala, Leon G J Tertoolen, Godfrey L Smith, Francis L Burton, Christine L Mummery

{"title":"Quantification of Muscle Contraction In Vitro and In Vivo Using MUSCLEMOTION Software: From Stem Cell-Derived Cardiomyocytes to Zebrafish and Human Hearts.","authors":"Berend J van Meer, Luca Sala, Leon G J Tertoolen, Godfrey L Smith, Francis L Burton, Christine L Mummery","doi":"10.1002/cphg.67","DOIUrl":"https://doi.org/10.1002/cphg.67","url":null,"abstract":"Quantification of contraction is essential to the study of cardiac diseases, injury, and responses to drugs. While there are many techniques to assess contractility, most rely on costly, dedicated hardware and advanced informatics, and can only be used in specific experimental models. We have developed an automated open-source software tool (MUSCLEMOTION) for use with standard imaging equipment, to assess contractility in vitro and in vivo and quantify responses to drugs and diseases. We describe high-speed and disturbance-free acquisition of images from either electrically paced or non-paced human pluripotent stem cell-derived cardiomyocytes, isolated adult cardiomyocytes, zebrafish hearts, and human echocardiograms. Recordings are then used as input for automated batch analysis by the MUSCLEMOTION software tool configured with specific settings and parameters tailored to the recording technique. Details on accuracy, interpretation, and troubleshooting are discussed. Acquisition duration depends on the experimental setup and aim, but quantification of drug or disease responses in an in vitro muscle model can typically be completed within a few hours. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":40007,"journal":{"name":"Current Protocols in Human Genetics","volume":" ","pages":"e67"},"PeriodicalIF":0.0,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cphg.67","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36523521","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: 13

Assessing Skewed X-Chromosome Inactivation 评估扭曲x染色体失活

Current Protocols in Human Genetics Pub Date : 2018-07-10 DOI: 10.1002/cphg.66

Thomas Liehr, Monika Ziegler, Sharon Löhmer, Anja Weise

引用次数: 2

Capture Hi-C Library Generation and Analysis to Detect Chromatin Interactions 捕获Hi-C库生成和分析检测染色质相互作用

Current Protocols in Human Genetics Pub Date : 2018-07-06 DOI: 10.1002/cphg.63

Giulia Orlando, Ben Kinnersley, Richard S. Houlston

{"title":"Capture Hi-C Library Generation and Analysis to Detect Chromatin Interactions","authors":"Giulia Orlando, Ben Kinnersley, Richard S. Houlston","doi":"10.1002/cphg.63","DOIUrl":"10.1002/cphg.63","url":null,"abstract":"Chromosome conformation capture (3C), coupled with next-generation sequencing (Hi-C), provides a means for deciphering not only the principles underlying genome folding and architecture, but more broadly, the role 3D chromatin structure plays in gene regulation and the replication and repair of DNA. The recently implemented modification, in situ Hi-C, maintains nuclear integrity during digestion and ligation steps, reducing random ligation of Hi-C fragments. Although Hi-C allows for genome-wide characterization of chromatin contacts, it requires high-depth sequencing to discover significant contacts. To address this, Capture Hi-C (CHi-C) enriches standard Hi-C libraries for regions of biological interest, for example by specifically targeting gene promoters, aiding identification of biologically significant chromatin interactions compared to conventional Hi-C, for an equivalent number of sequence reads. Illustrating the application of CHi-C applied to genome-wide analysis of chromatin interactions with promoters, we detail the protocols for in situ Hi-C and CHi-C library generation for sequencing, as well as the bioinformatics tools for data analysis. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":40007,"journal":{"name":"Current Protocols in Human Genetics","volume":"98 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cphg.63","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10645056","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: 10

Reporting of Clinical Genome Sequencing Results 临床基因组测序结果报告

Current Protocols in Human Genetics Pub Date : 2018-07-06 DOI: 10.1002/cphg.61

Cui Song, Hatice Duzkale, Jun Shen

引用次数: 1