Pubblicazione scientifica

Fluorescence In situ Hybridization: Cell-Based Genetic Diagnostic and Research Applications

FILONE TECNOLOGICO	TEMA
trasformazione e commercializzazione

RIASSUNTO

Fluorescence in situ hybridization (FISH) is a macromolecule recognition technology based on the complementary nature of DNA or DNA/RNA double strands. Selected DNA strands incorporated with fluorophore-coupled nucleotides can be used as probes to hybridize onto the complementary sequences in tested cells and tissues and then visualized through a fluorescence microscope or an imaging system. This technology was initially developed as a physical mapping tool to delineate genes within chromosomes. Its high analytical resolution to a single gene level and high sensitivity and specificity enabled an immediate application for genetic diagnosis of constitutional common aneuploidies, microdeletion/microduplication syndromes, and subtelomeric rearrangements. FISH tests using panels of gene-specific probes for somatic recurrent losses, gains, and translocations have been routinely applied for hematologic and solid tumors and are one of the fastest-growing areas in cancer diagnosis. FISH has also been used to detect infectious microbias and parasites like malaria in human blood cells. Recent advances in FISH technology involve various methods for improving probe labeling efficiency and the use of super resolution imaging systems for direct visualization of intra-nuclear chromosomal organization and profiling of RNA transcription in single cells. Cas9-mediated FISH (CASFISH) allowed in situ labeling of repetitive sequences and single-copy sequences without the disruption of nuclear genomic organization in fixed or living cells. Using oligopaint-FISH and super-resolution imaging enabled in situ visualization of chromosome haplotypes from differentially specified single-nucleotide polymorphism loci. Single molecule RNA FISH (smRNA-FISH) using combinatorial labeling or sequential barcoding by multiple round of hybridization were applied to measure mRNA expression of multiple genes within single cells. Research applications of these single molecule single cells DNA and RNA FISH techniques have visualized intra-nuclear genomic structure and sub-cellular transcriptional dynamics of many genes and revealed their functions in various biological processes.

DATA

Data di pubblicazione: 05/09/2016

AUTORI

CUI CHENGHUA
SHU WEI
LI PEINING

ENTE DI AFFERENZA

LAB OF CLINICAL CYTOGENETICS DEPT GENETICS YALE SCHOOL OF MEDICINE NEW HAVEN CT USA
DEPT PATHOLOGY INST HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACAD MEDICAL SCIENCES TIANJIN CHINA
DEPT CELL BIOLOGY & GENETICS GUANGXI MEDICAL UNIV NANNING CHINA

RIVISTA

Frontiers in Cell and Developmental Biology, 4, 89

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