Current Protocols in Human Genetics This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.

Molecular Analysis of Genetic Markers for Non-Hodgkin Lymphomas.
Authors: Sholl LM, Longtine J, Kuo FC Abstract Molecular analysis complements the clinical and histopathologic tools used to diagnose and subclassify hematologic malignancies. The presence of clonal antigen-receptor gene rearrangements can help to confirm the diagnosis of a B or T cell lymphoma and can serve as a fingerprint of that neoplasm to be used in identifying concurrent disease at disparate sites or recurrence at future time points. Certain lymphoid malignancies harbor a characteristic chromosomal translocation, a finding that may have significant implications for an individual's prognosis or respo...
Source: Current Protocols in Human Genetics - April 7, 2017 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Overview of Genetic Diagnosis in Cancer.
Authors: Korf BR, Mikhail FM Abstract Both cytogenetic and molecular genetic studies can contribute to the management of patients with cancer. In some cases, genetic markers are specific to particular tumor types and are useful in diagnosis. This can be helpful in distinguishing histologically similar tumors that may respond differently to treatment and can sometimes be of prognostic value. Genetic markers can also be tools for following the response of a tumor to therapy, providing a sensitive means to detect relapse. This introductory unit considers some of the types of genetic changes that occur in asso...
Source: Current Protocols in Human Genetics - April 7, 2017 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Computational Approach to Measuring Myocyte Disarray in Animal Models of Heart Disease.
Authors: Wan W, Leinwand L Abstract In cardiovascular disease research, studies often include measuring cardiac function and performing histological examination of heart tissue. After measuring contractility, hearts from animals such as mice and rats are often frozen or fixed, sliced, and stained to quantify the morphology of various structures such as extracellular matrix proteins, cell nuclei, and F-actin. Traditional scoring methods have largely consisted of assessing sections of images for the presence or absence of myocyte disarray. These approaches require unbiased manual assessment, which can requir...
Source: Current Protocols in Human Genetics - April 7, 2017 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Diagnosis of Spinocerebellar Ataxias Caused by Trinucleotide Repeat Expansions.
Authors: Martindale JE Abstract Spinocerebellar ataxias (SCAs) are a group of disorders that are both clinically and genetically heterogeneous. They usually demonstrate onset in adulthood, but some forms may have juvenile or infantile onset. There are many different types of SCA, demonstrating different modes of inheritance and types of mutation. The most common forms are due to dominantly inherited expansions in trinucleotide repeat sequences located within the coding region of the relevant genes, and these are readily identifiable by molecular genetic testing. In general, it is possible to test for these...
Source: Current Protocols in Human Genetics - January 12, 2017 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Human Induced Pluripotent Stem (hiPS) Cells from Urine Samples: A Non-Integrative and Feeder-Free Reprogramming Strategy.
Authors: Steichen C, Si-Tayeb K, Wulkan F, Crestani T, Rosas G, Dariolli R, Pereira AC, Krieger JE 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" repr...
Source: Current Protocols in Human Genetics - January 12, 2017 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Detecting APC Gene Mutations in Familial Adenomatous Polyposis (FAP).
Authors: Nallamilli BR, Hegde M 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 ...
Source: Current Protocols in Human Genetics - January 12, 2017 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Assay for Transposase-Accessible Chromatin Using Sequencing (ATAC-seq) Data Analysis.
Authors: Miskimen KL, Chan ER, Haines JL Abstract The study of epigenetic properties of the human genome, including structural modifications of DNA and chromatin, has increased tremendously as mounting evidence has demonstrated how much epigenetics affects human gene expression. Buenrostro et al. have developed a rapid method, requiring low numbers of living cells as input, for examining chromatin accessibility across the epigenome, known as the assay for transposase-accessible chromatin using sequencing (ATAC-seq). The overall goal of this unit is to provide a thorough ATAC-seq data analysis plan, as wel...
Source: Current Protocols in Human Genetics - January 12, 2017 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Culturing and Neuronal Differentiation of Human Dental Pulp Stem Cells.
Authors: Goorha S, Reiter LT Abstract A major issue in studying human neurogenetic disorders, especially rare syndromes affecting the nervous system, is the ability to grow neuronal cultures that accurately represent these disorders for analysis. Although there has been some success in generating induced pluripotent stem (iPS) cells from both skin and blood, there are still limitations to the collection and production of iPS cells from these biospecimens. We have had significant success in collecting and growing human dental pulp stem (DPS) cells from exfoliated teeth sent to our laboratory by the parents ...
Source: Current Protocols in Human Genetics - January 12, 2017 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Highly Expandable Human iPS Cell-Derived Neural Progenitor Cells (NPC) and Neurons for Central Nervous System Disease Modeling and High-Throughput Screening.
Authors: Cheng C, Fass DM, Folz-Donahue K, MacDonald ME, Haggarty SJ 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 expandab...
Source: Current Protocols in Human Genetics - January 12, 2017 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Molecular Analysis of Gene Rearrangements and Mutations in Acute Leukemias and Myeloid Neoplasms.
Authors: Sholl LM, Longtine J, Kuo FC Abstract A subset of acute leukemias and other myeloid neoplasms contains specific genetic alterations, many of which are associated with unique clinical and pathologic features. These alterations include chromosomal rearrangements leading to oncogenic fusion proteins or alteration of gene expression by juxtaposing oncogenes to enhancer elements, as well as mutations leading to aberrant activation of a variety of proteins critical to hematopoietic progenitor cell proliferation and differentiation. Molecular analysis is central to diagnosis and clinical management of le...
Source: Current Protocols in Human Genetics - January 12, 2017 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Generating Exome Enriched Sequencing Libraries from Formalin-Fixed, Paraffin-Embedded Tissue DNA for Next-Generation Sequencing.
Authors: Marosy BA, Craig BD, Hetrick KN, Witmer PD, Ling H, Griffith SM, Myers B, Ostrander EA, Stanford JL, Brody LC, Doheny KF Abstract This unit describes a technique for generating exome-enriched sequencing libraries using DNA extracted from formalin-fixed paraffin-embedded (FFPE) samples. Utilizing commercially available kits, we present a low-input FFPE workflow starting with 50 ng of DNA. This procedure includes a repair step to address damage caused by FFPE preservation that improves sequence quality. Subsequently, libraries undergo an in-solution-targeted selection for exons, followed by sequenci...
Source: Current Protocols in Human Genetics - January 12, 2017 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

High-Risk Screening of Fabry Disease: Analysis of Fifteen Urinary Methylated and Non-Methylated Gb3 Isoforms Using Tandem Mass Spectrometry.
Authors: Abaoui M, Boutin M, Lavoie P, Auray-Blais C Abstract Fabry disease is a multisystemic, X-linked lysosomal storage disorder caused by mutations in the GLA gene, leading to α-galactosidase A deficiency and resulting in the accumulation of glycosphingolipids in different tissues and biological fluids. Glycosphingolipid biomarkers, such as globotriaosylceramide (Gb3 ) isoforms, globotriaosylsphingosine (lyso-Gb3 ) and related analogs, and galabiosylceramide (Ga2 ) isoforms and analogs, are found to be abnormally increased in urine and in plasma of Fabry patients and have the potential to be used as s...
Source: Current Protocols in Human Genetics - October 13, 2016 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Pronuclear Injection-Based Targeted Transgenesis.
Authors: Schilit SL, Ohtsuka M, Quadros RM, Gurumurthy CB Abstract Microinjection of DNA expression cassettes into fertilized zygotes has been a standard method for generating transgenic animal models. While efficient, the injected DNA integrates randomly into the genome, leading to potential disruption of endogenous genes or regulatory elements, variation in copy number, or integration into heterochromatic regions that inhibit transgene expression. A recently developed method addresses such pitfalls of traditional transgenesis by targeting the transgene to predetermined sites in the genome that can safely...
Source: Current Protocols in Human Genetics - October 13, 2016 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Acylglycine Analysis by Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS).
Authors: Hobert JA, Liu A, Pasquali M Abstract Quantitative analysis of urine acylglycines has shown to be a highly sensitive and specific method with proven clinical utility for the diagnosis of several inherited metabolic disorders including: medium chain acyl-CoA dehydrogenase deficiency, multiple acyl-CoA dehydrogenase deficiency, short chain acyl-CoA dehydrogenase deficiency, 3-methylcrotonyl-CoA carboxylase deficiency, 2-methylbutyryl-CoA dehydrogenase deficiency, isovaleric acidemia, propionic academia, and isobutyryl-CoA dehydrogenase deficiency. Here, a method that is currently performed using ult...
Source: Current Protocols in Human Genetics - October 13, 2016 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research

Molecular Diagnosis of Myotonic Dystrophy.
Authors: Chakraborty S, Vatta M, Bachinski LL, Krahe R, Dlouhy S, Bai S Abstract Myotonic dystrophy types 1 (DM1) and 2 (DM2) are autosomal dominant, microsatellite repeat expansion disorders that affect muscle function. Myotonic dystrophy type 1 is caused by CTG repeat expansion in the 3' UTR region of the DMPK gene. Patients with DM2 have expansion of CCTG repeats in intron 1 of the CNBP gene. In this unit, we review and discuss the clinical phenotypes, genetic mutations causing the diseases, and the molecular diagnostic approaches and tools that are used to determine repeat sizes in DM1/2. In summary, t...
Source: Current Protocols in Human Genetics - October 13, 2016 Category: Genetics & Stem Cells Tags: Curr Protoc Hum Genet Source Type: research