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Gene editing using the CRISPR/Cas9 system is an extremely efficient approach for generating mutations within the genomic DNA of immortalized cell lines. This procedure begins with a straightforward cloning step to generate a single plasmid encoding the Cas9 enzyme as well as a synthetic guide RNA (sgRNA)which is selected to target specific sites within the genome. This plasmid is transfected into cells either alone, in order to generate random insertion-deletion alleles ("indels") at the desired locus via the nonhomologous end-joining pathway, or in conjunction with a homology-directed repair template oligonucleotide to generate a specific point mutation. Here we describe a procedure to perform gene editing in IMCD3 andHEK293 cells and to subsequently isolate clonal cell lines carrying mutations of interest.
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MicroRNAs (miRNAs) are small regulatory RNAs that play a crucial role in posttranscriptional gene regulation. Over two thousand miRNAs have been identified in humans, and many of them are conserved in other species. miRNAs are implicated in fundamental cellular functions, including development and disease. In the last decade, there has been an overwhelming amount of data contributing to the understanding of miRNA biogenesis and their target genes. Moreover, a significant amount of work has been carried out in developing miRNA biomarkers and therapeutics for various disease conditions. RNA-based markers and therapeutics have been proven to have a clinical impact, and many of these miRNA-based therapies are at various stages of human clinical trials and clinical applications. Notably, miRNAs are also found in exosomes, and are considered to impart intercellular communication and function via several different modalities, including tunneling nanotubes. In spite of our understanding of miRNA biology and function, there are many challenges in effectively using miRNAs as biomarkers and therapeutic agents in clinical applications. In this Special Issue, we are inviting reviews, perspectives, and original research articles to address some of these challenges. Topics will include, but are not limited to, miRNA biogenesis, clinical applications, extracellular function, biomarkers, miRNA immune regulation, signaling pathways, and preclinical models.
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Cancer is a multifaceted disease in which genetic changes induce uncontrolled tumor growth. Genomic characterization of cancer is now leading to better diagnostic, prognostic and predictive biomarkers, and effective individualized management. 'Fast Facts: Comprehensive Genomic Profiling' provides a crash course in the science, methods and application of genomic profiling. Assuming only the most basic knowledge - or memory - of cell biology, the authors provide an overview of DNA and RNA biology and next-generation sequencing. This sets in context the descriptions of prognostic and predictive biomarkers for different cancer types and genomic-based treatments. Finally, but importantly, some of the practicalities of gaining and interpreting genomic information are described. Whether you need a primer or a refresher, this short colorful book demystifies this complex subject. Contents: • Genetic mutations and biomarkers • Understanding next-generation sequencing • Elements of comprehensive genomic profiles • Role in precision oncology • Predictive and prognostic biomarkers • Overcoming barriers to genotype-directed therapy
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MicroRNAs (miRNAs) are small regulatory RNAs that play a crucial role in posttranscriptional gene regulation. Over two thousand miRNAs have been identified in humans, and many of them are conserved in other species. miRNAs are implicated in fundamental cellular functions, including development and disease. In the last decade, there has been an overwhelming amount of data contributing to the understanding of miRNA biogenesis and their target genes. Moreover, a significant amount of work has been carried out in developing miRNA biomarkers and therapeutics for various disease conditions. RNA-based markers and therapeutics have been proven to have a clinical impact, and many of these miRNA-based therapies are at various stages of human clinical trials and clinical applications. Notably, miRNAs are also found in exosomes, and are considered to impart intercellular communication and function via several different modalities, including tunneling nanotubes. In spite of our understanding of miRNA biology and function, there are many challenges in effectively using miRNAs as biomarkers and therapeutic agents in clinical applications. In this Special Issue, we are inviting reviews, perspectives, and original research articles to address some of these challenges. Topics will include, but are not limited to, miRNA biogenesis, clinical applications, extracellular function, biomarkers, miRNA immune regulation, signaling pathways, and preclinical models.
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MicroRNAs (miRNAs) are small regulatory RNAs that play a crucial role in posttranscriptional gene regulation. Over two thousand miRNAs have been identified in humans, and many of them are conserved in other species. miRNAs are implicated in fundamental cellular functions, including development and disease. In the last decade, there has been an overwhelming amount of data contributing to the understanding of miRNA biogenesis and their target genes. Moreover, a significant amount of work has been carried out in developing miRNA biomarkers and therapeutics for various disease conditions. RNA-based markers and therapeutics have been proven to have a clinical impact, and many of these miRNA-based therapies are at various stages of human clinical trials and clinical applications. Notably, miRNAs are also found in exosomes, and are considered to impart intercellular communication and function via several different modalities, including tunneling nanotubes. In spite of our understanding of miRNA biology and function, there are many challenges in effectively using miRNAs as biomarkers and therapeutic agents in clinical applications. In this Special Issue, we are inviting reviews, perspectives, and original research articles to address some of these challenges. Topics will include, but are not limited to, miRNA biogenesis, clinical applications, extracellular function, biomarkers, miRNA immune regulation, signaling pathways, and preclinical models.
Choose an application
Gene editing using the CRISPR/Cas9 system is an extremely efficient approach for generating mutations within the genomic DNA of immortalized cell lines. This procedure begins with a straightforward cloning step to generate a single plasmid encoding the Cas9 enzyme as well as a synthetic guide RNA (sgRNA) which is selected to target specific sites within the genome. This plasmid is transfected into cells either alone, in order to generate random insertion-deletion alleles ("indels") at the desired locus via the nonhomologous end-joining pathway, or in conjunction with a homology-directed repair template oligonucleotide to generate a specific point mutation. Here we describe a procedure to perform gene editing in IMCD3 and HEK293 cells and to subsequently isolate clonal cell lines carrying mutations of interest.
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Genetics --- Human genetics. --- Genetics, Medical
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Handbook of Clinical Adult Genetics and Genomics: A Practice-Based Approach provides a thorough overview of genetic disorders that are commonly encountered in adult populations and supports the full translation of adult genetic and genomic modalities into clinical practice. Expert chapter authors supplement foundational knowledge with case-based strategies for the evaluation and management of genetic disorders in each organ system and specialty area. Topics discussed include employing genetic testing technologies, reporting test results, genetic counseling for adult patients, medical genetics referrals, issues of complex inheritance, gene therapy, and diagnostic and treatment criteria for developmental, cardiovascular, gastrointestinal, neuropsychiatric, pulmonary issues, and much more. Employs clinical case studies to demonstrate how to evaluate, diagnosis and treat adult patients with genetic disorders Offers a practical framework for establishing an adult genetics clinic, addressing infrastructure, billing, counseling, and challenges unique to adult clinical genetics Features chapter contributions from authors at leading adult genetics institutions in the US and abroad --
Genetics --- Human genetics --- Medical genetics --- Genetics, Medical
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