This volume presents a list of cutting-edge protocols for the study of CRISPR-Cas defense systems and their applications at the genomic, genetic, biochemical and structural levels. CRISPR: Methods and Protocols guides readers through techniques that have been developed specifically for the analysis of CRISPR-Cas and techniques adapted from standard protocols of DNA, RNA and protein biology. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, CRISPR: Methods and Protocols provides a broad list of tools and techniques to study the interdisciplinary aspects of the prokaryotic CRISPR-Cas defense systems.

This book encompasses major progress and future directions in cytochrome P450 (P450) research. Included are contributions by pioneers in the discovery of P450, with chapters on the molecular and functional properties of P450 and cutting-edge applications knowledge from various fields. P450 research has its roots in metabolism, but the true beginning was in 1962 with the publication by Tsuneo Omura and Ryo Sato in "The Journal of Biological Chemistry "on their discovery of the cytochrome." "Following this groundbreaking study, over the last half-century, research has revealed that many forms of P450 exist in animals, plants and microorganisms. P450 research has expanded into many different fields including medicine, agriculture and biotechnology and has drawn the attention of industries for its bioengineering applications, such as drug development and creation of the blue rose . Also, research on nuclear receptors, which has grown out of research on the regulatory mechanisms of P450 genes, has become an important area in biology, medical science, pharmacology and clinical medicine for example, with recent developments in personalized medicines. This book will draw readers into the important and exciting world of P450 and will encourage young students and scientists in P450 research to continue expanding the field via new approaches."

This stimulating analysis reviews the broad potential of animal models to foster a deeper understanding of human pathology, strengthen connections between genetic and behavioral studies, and develop more effective treatments for mental disorders. Widely-studied and lesser-used species are examined in models that capture features along the continuum of normative and pathological behavior. The models highlight genetic causes of core features, or endophenotypes, of developmental, internalizing, and externalizing disorders, as well as dementia. Expert contributors address questions ranging from how suitable species are chosen for study to the costs and benefits of using inbred versus outbred strains, and the effects of housing environment on subject animals. Larger issues addressed include how to evaluate the applicability of animal behavioral models to the human condition and how these models can harness emerging molecular technologies to further our understanding of the genetic basis of mental illness. Included in the coverage: Mating and fighting in Drosophila. Attachment and social bonding. Impulsivity in rodents and humans. Animal models of cognitive decline. Animal models of social cognition. Future directions for animal models in behavioral genetics. A detailed map of where this evolving field is headed, Animal Models of Behavior Genetics shows geneticists, molecular biologists, and cognitive neuroscientists paths beyond established concepts toward a more knowledgeable and collaborative future.

Using both epidemiological and model organism approaches, we have gained new insights into the physiological and molecular aspects of aging, which has led to significant advancements in potential anti-aging strategies. Reviews on Biomarker Studies in Aging and Anti-Aging Research presents a series of reviews in various aspects of aging and age-related disease research along with several methods which have shown progress as potential anti-aging approaches. The book is aimed at researchers in the areas of aging and chronic disease, as well as to clinical scientists, physicians and major drug companies. It provides important information on disease mechanisms, and each chapter is presented in the context of the aging process, specific chronic diseases or different therapeutic areas.

This books aims to provide an up-to-date guide to using recombinases in the laboratory. Chapters guide the reader through construction of transgenic animals, recombinase-mediated cassette exchange (RMCE), spatial control of recombinase expression and in vivo delivery approaches, immunohistochemistry and fluorescence-based phenotyping methods, biochemical methods, and recombinase-based production of minimal DNA vectors. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Site-Specific Recombinases: Methods and Protocols aims to serve as a valuable resource for working with recombinases, and inspires further creativity in the field.

This book provides a review of the multitude of nucleic acid polymerases, including DNA and RNA polymerases from Archea, Bacteria and Eukaryota, mitochondrial and viral polymerases, and other specialized polymerases such as telomerase, template-independent terminal nucleotidyl transferase and RNA self-replication ribozyme. Although many books cover several different types of polymerases, no book so far has attempted to catalog all nucleic acid polymerases. The goal of this book is to be the top reference work for postgraduate students, postdocs, and principle investigators who study polymerases of all varieties. In other words, this book is for polymerase fans by polymerase fans.

Nucleic acid polymerases play a fundamental role in genome replication, maintenance, gene expression and regulation. Throughout evolution these enzymes have been pivotal in transforming life towards RNA self-replicating systems as well as into more stable DNA genomes. These enzymes are generally extremely efficient and accurate in RNA transcription and DNA replication and share common kinetic and structural features. How catalysis can be so amazingly fast without loss of specificity is a question that has intrigued researchers for over 60 years. Certain specialized polymerases that play a critical role in cellular metabolism are used for diverse biotechnological applications and are therefore an essential tool for research.

This book presents an investigation of language contact, focusing on Northwestern China. It breaks down the barrier between human sciences and natural sciences in order to reconsider the diversity of languages on the basis of the latest research findings from genetics, linguistics, and other domains, offering valuable insights into when and how the divergence of languages and genes began and language and gene admixture and replacement occurred. The book focuses on language evolution between the border of Gansu and Qinghai Province in China, but the research doesn't neglect the area beyond China's northern borders. Manchu, a dying language belonging to the Tungusic group, is also studied to enhance our understanding of language replacement. This work is the result of a four-year collaboration between teams of geneticists and linguists in France and China.

PIWI-interacting RNAs (piRNAs) are the third and most-recently discovered group of silencing-inducible small RNAs in animals. PIWI-Interacting RNAs: Methods and Protocols provides the most recent methods and protocols for studying piRNAs in the gonads of a wide range of species, as well as in any other organs where piRNAs may be detected. Comprehensive high-throughput sequencing analysis of piRNAs in embryos, testes and ovaries of D. melanogaster, as well as in mouse and rat testes, has raised the profile of piRNAs and thus further accelerated piRNA studies. In addition to C. elegans, other model species such as Drosophila melanogaster, Arabidopsis thaliana and mice, along with cultured cell lines such as HeLa and Drosophila Schneider 2 (S2), and other such organisms have been used to address the fundamental questions of the biogenesis and functions of RNAi-triggering small non-coding RNAs. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, PIWI-Interacting RNAs: Methods and Protocols seeks to serve both established researchers and newcomers to the field to progress towards the ultimate goal of understanding the mechanisms and actions of piRNAs.

Lynch syndrome (LS) is the most common cause of inherited colorectal cancer, a disease with a high mortality rate. An estimated 37,000 of diagnosed colorectal cancer cases worldwide are attributed to Lynch syndrome each year. Intensive cancer screening, with early initiation and frequent follow-up, can reduce colorectal cancer incidence and mortality in LS patients. This book provides an up-to-date overview on the genetic and epigenetic basis of Lynch syndrome. It evaluates clinical features of the disease and critically comments on molecular tools available for identifying mutations responsible for Lynch syndrome; in addition the importance of functional assays that can help clarify the clinical nature of identified mutations is also discussed. The book also focuses on challenges in genetic counselling of at-risk individuals and discusses related ethical issues. The purpose of the book is to give a concise knowledge base for the broader scientific and medical community, including genetic counselors, in order to improve awareness on the potential impact that the diagnosis of LS has on treatment, management and surveillance of LS patients.

This volume presents the latest protocols for both laboratory and bioinformatics based analyses in the field of marine genomics. The chapters presented in the book cover a wide range of topics, including the sampling and genomics of bacterial communities, DNA extraction in marine organisms, high-throughput sequencing of whole mitochondrial genomes, phylogenomics, SNP discovery, SNP-arrays for species identification, digital PCR-based quantification methods, environment DNA for invasive species surveillance and monitoring, microarrays for the detection of waterborne pathogens, DNA barcoding of marine biodiversity, metabarcoding protocols for marine eukaryotes, analytical protocols for the visualization of eukaryotic diversity, and applications for genomic data to benthic indices for environmental monitoring. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Marine Genomics: Methods and Protocols is a valuable resource for researchers, students, and policy makers in the field of marine biology.

The concept of immunotherapy was in infancy when the first edition was written; since then, major advances have been made, not only with several prominent clinical trials, but also with the approval of cell-based therapy by the FDA for the treatment of cancer in 2010. These events resulted in a gradually narrowing gap between early scientific knowledge and the late development of immune-based therapies. Consequently, the significance and magnitude of these advances warranted a revision of this contribution; this revised edition will provide a deeper understanding of the recent advances and discoveries related to the function of the immune response and their applications in the development of novel therapies to treat human diseases. Some of the key discoveries during the past five years include: the identification of the new subsets of helper T cells; new cytokines and their networks; and novel signal transduction mechanisms. For example, the identification of TH17 subset of helper T cells, in addition to TH1 and TH2 cells, not only advanced our understanding of the function of the basic immune response, but also raised our awareness of the possible etiology and pathogenesis of diseases such as allergy, asthma, rheumatoid arthritis, and other auto-immune/immune system based diseases. The newly identified powerful cytokine networks, that regulate both innate and acquired immune responses, emerged as a result of the finding of new cell types such as innate lymphoid cells and iNKT. Identification of the novel cytokines and their networks has advanced our knowledge of the mechanisms involved in the maintenance of tissue homeostasis, including inflammation and tissue repair during stress and injury. The development of HIV vaccines has also seen dramatic changes over the last few years. There has been a shift from a sole focus on T cell vaccines to a holistic approach that pertains to the induction of both humoral and cellular elements. This entails the induction of antibodies - both binding and neutralizing - to prevent infection. The cellular vaccination produces a safety net of CD8+ T-cell responses to suppress the replication of the virus in the infected patients, and both of the effector arms are aided by helper T cells. From the perspective of clinical applications, significant advances have also been made in: oral immunotherapy for allergic disease, the possible treatment of HIV infection, the development of new monoclonal antibodies and their fragments to treat human diseases, and immune cell based therapies for cancer.

This book focus on genetic diagnostics for Uniparental Disomy (UPD), a chromosomal disorder defined by the exceptional presence of a chromosome pair derived from only one parent, which leads to a group of rare diseases in humans. First the molecular and cytogenetic background of UPD is described in detail; subsequently, all available information of the various chromosomal origins and the latest findings on genotype-phenotype correlations and clinical consequences are discussed. Numerous personal reports from families with a child suffering from a UPD-induced syndrome serve to complement the scientific and clinical aspects. Their experiences with genetic counseling and living with a family member affected by this chromosomal aberration present a vivid picture of what UPD means for its victims.

This book provides information on a wide variety of issues ranging from genetics to clinical description of the syndromes, genetic testing and counseling, and clinical management including surveillance, surgical and prophylactic interventions, and chemoprevention. Moreover, current hot issues, such as the identification of novel causal genes and the challenges we face, and the relevance of cancer risk modifiers, both genetic and environmental, are also discussed. This reference book is great for geneticists, oncologists, genetic counselors, researchers, clinicians, surgeons and nurses dedicated to, or interested in, hereditary cancer. The best and most recognized experts in the field have contributed to this project, guaranteeing updated information, accuracy and the discussion of topical issues.

In vitro mutagenesis remains a critical experimental approach for investigating gene and protein function at the cellular level. This volume provides a wide variety of updated and novel approaches for performing in vitro mutagenesis using such methods as genome editing, transposon (Tn) mutagenesis, site-directed, and random mutagenesis. In Vitro Mutagenesis: Methods and Protocols guides readers through methods for gene and genome editing, practical bioinformatics approaches for identifying mutagenesis targets, and novel site-directed and random mutagenesis approaches aimed at gaining a better understanding of protein-protein and protein-cofactor interactions. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, In Vitro Mutagenesis: Methods and Protocols aims to provide a highly accessible and practical manual for current and future molecular biology researchers, from the beginner practitioner to the advanced investigator in fields such as molecular genetics, biochemistry, and biochemical and metabolic engineering.

This volume provides broad coverage of computational and mathematical techniques and concepts related to the field of comparative genomics. The topics covered in the chapters range from those concerned with general techniques and concepts that apply to all organisms to others that are more specialized, covering specific biological systems such as viruses, Drosophila, and Homo sapiens. Written in the highly successful Methods in Molecular Biology series format, by authors who are active researchers in the field, many chapters include step-by-step procedures, which illustrate practical applications of the techniques described. Cutting-edge and thorough, Comparative Genomics: Methods and Protocols should be useful to students and researchers in the continually growing and exciting field of comparative genomics.

This volume assembles a broad spectrum of methods used in long non-coding RNAs (lncRNA) research, ranging from computational annotation of lncRNA genes to molecular and cellular analyses of the function of individual lncRNA. Long Non-Coding RNAs: Methods and Protocols also discusses methods used to study circular RNAs and RNA splicing, as well as influential findings on lncRNA in human diseases. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Thorough and cutting-edge, Long Non-Coding RNAs: Methods and Protocols is a must-have for molecular biologists, cell and developmental biologists, specialists who conduct disease-oriented research, and bioinformatics experts who seek a better understanding on lncRNA expression and function by computational analysis of the massive sequencing data that are rapidly accumulating in recent years.

This book presents the state of the art of type 2 diabetes genetics, from the process of genetic discovery to its interpretation and clinical application, and illustrates a model for other complex human phenotypes.The first section explores genome-wide association studies, the extension of this method to less accessible phenotypes and the arrival of next-generation sequencing. A further section goes beyond genetics to illustrate how other data sources can help interpret genetic data, such as leveraging population diversity, the correlation of genetic associations with physiological measurements, gene expression modulation, environmental factors and our microbial commensals. The third section describes advances in elucidating the complex path from association to function using in-depth sequencing and functional studies of the cellular and molecular effects of genes in the loci identified by genetics. The final section links our current understanding with clinically relevant questions, such as prediction, interactions with drugs or nutrients, and disease prevention, and paints a realistic but hopeful vision of the future.

The primary purpose of this book and its companion volume The Behavioral Genetics of Nicotine and Tobacco is to explore the ways in which recent studies on nicotine and its role in tobacco addiction have opened our eyes to the psychopharmacological properties of this unique and fascinating drug. While The Behavioral Genetics of Nicotine and Tobacco considers the molecular and genetic factors which influence behavioral responses to nicotine and how these may impact on the role of nicotine in tobacco dependence, the present book focuses on the complex neural and psychological mechanisms that mediate nicotine dependence in experimental animal models and their relationship to tobacco addiction in humans. These volumes will provide readers a contemporary overview of current research on nicotine psychopharmacology and its role in tobacco dependence from leaders in this field of researchand will hopefully prove valuable to those who are developing their own research programmes in this important topic.

This stimulating volume uses multiple lenses to analyze the complex causes of health disparities affecting minorities, in particular African Americans, and explains how this knowledge can be used to reduce their destructive effects. Pinpointing genetic, non-genetic, and epigenetic factors underlying health conditions common to the population-including heart disease, hypertension, diabetes, and cancer-the author traces intricate links among these factors in the current environmental and social context. The section on non-genetic factors in health disparities, such as social determinants and health behaviors, adds depth to the ongoing discourse on public health and health policy objectives. And the chapters on gene/environment interactions outline the vast potential for developing new multidisciplinary frontiers in shrinking health inequities and personalizing care. Included in the coverage: The African diaspora and disease-specific disparities The genetic basis to health disparities The role of epigenetics Economic factors and health Psychological issues and how they affect disparities Gene-environment interactions in health disparities Race, a biological or social concept Compelling and accessible, Health Outcomes in a Foreign Land will challenge and inspire medical students, epidemiologists, public health professionals, biomedical research scientists, and social scientists to go farther in their work. A wider audience would include policymakers, government officials, nurses, physicians, lawyers, economists, community outreach investigators, and interested general readers.

This second edition volume expands on the previous edition by presenting updated protocols for several of the techniques described in the first edition of Drosophila: Methods and Protocols and current methods that cover recent breakthroughs in Drosophila research. The book begins with a description of FlyBase--a database of genes and genomes--followed by the presentation of systems for versatile gene expression in the fly. The first few chapters in this book detail gene knockdown and editing, including CRISPR-Cas9 and protein knockdown. The next few chapters are devoted to methods describing live imaging of different tissues and organs, followed by chapters on how to quantify image data and how to probe tissue mechanics by laser ablation. The next two chapters provide methods for analyzing transcription followed by protocols to study growth, metabolism, ageing, and behavior in Drosophila. This volume concludes with chapters on electrophysiological recordings and methods to establish cell lines. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Drosophila: Methods and Protocols, Second Edition is a valuable source of hands-on protocols and reviews for molecular, cell, and developmental biologists using Drosophila as model systems in their work.

This book presents an overview of the RNA networks controlling gene expression in fungi highlighting the remaining questions and future challenges in this area.

It covers several aspects of the RNA-mediated mechanisms that regulate gene expression in model yeasts and filamentous fungi, organisms of great importance for industry, medicine and agriculture. It is estimated that there are more than one million fungal species on the Earth. Despite their diversity (saprophytic, parasitic and mutualistic), fungi share common features distinctive from plants and animals and have been grouped taxonomically as an independent eukaryotic kingdom. In this book, 15 chapters written by experts in their fields cover the RNA-dependent processes that take place in a fungal cell ranging from formation of coding and non-coding RNAs to mRNA translation, ribosomal RNA biogenesis, gene silencing, RNA editing and epigenetic regulation.

Current knowledge of the mechanisms that regulate DNA repair has grown significantly over the past years with technology advances such as RNA interference, advanced proteomics and microscopy as well as high throughput screens. The third edition of DNA Repair Protocols covers various aspects of the eukaryotic response to genomic insult including recent advanced protocols as well as standard techniques used in the field of DNA repair. Both mammalian and non-mammalian model organisms are covered in the book, and many of the techniques can be applied with only minor modifications to other systems than the one described. Written in the highly successful Methods in Molecular Biology? series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory. Thorough and intuitive, DNA Repair Protocols, Third Edition provides expert guidance for DNA repair, recombination, and replication.

Ribonucleic acid (RNA) binding proteins currently number in the thousands and defects in their function are at the heart of diseases such as cancer and neurodegeneration. RNA binding proteins have become implicated in the intricate control of surprisingly diverse biological settings, such as circadian rhythm, stem cell self-renewal, oncogenesis and germ cell development. This book surveys a range of genome-wide and systems approaches to studying RNA binding proteins, the importance of RNA binding proteins in development, cancer and circadian rhythm.