Pubblicazione scientifica

Three-dimensional analysis of single molecule FISH in human colon organoids

FILONE TECNOLOGICO	TEMA
trasformazione e commercializzazione

RIASSUNTO

ABSTRACT

The culturing of mini-organs (organoids) in three-dimensions (3D) presents a simple and powerful tool to investigate the principles underlying human organ development and tissue self-organization in both healthy and diseased states. Applications of single molecule analysis are highly informative for a comprehensive understanding of the complexity underlying tissue and organ physiology. To fully exploit the potential of single molecule technologies, the adjustment of protocols and tools to 3D tissue culture is required. Single molecule RNA fluorescence in situ hybridization (smFISH) is a robust technique for visualizing and quantifying individual transcripts. In addition, smFISH can be employed to study splice variants, fusion transcripts as well as transcripts of multiple genes at the same time. Here, we develop a 3-day protocol and validation method to perform smFISH in 3D in whole human organoids. We provide a number of applications to exemplify the diverse possibilities for the simultaneous detection of distinct mRNA transcripts, evaluation of their spatial distribution and the identification of divergent cell lineages in 3D in organoids.

DATA

Data di pubblicazione: 30/07/2019

AUTORI

OMERZU MANJA
FENDERICO NICOLA
DE BARBANSON BUYS
SPRANGERS JOEP
DE RIDDER JEROEN
MAURICE MADELON M

ENTE DI AFFERENZA

ONCODE INST & DEPT CELL BIOLOGY CTR MOLECULAR MEDICINE UNIV MEDICAL CTR UTRECHT 3584 CX UTRECHT NETHERLANDS
ONCODE INST & DEPT GENETICS CTR MOLECULAR MEDICINE UNIV MEDICAL CTR UTRECHT 3584 CG UTRECHT NETHERLANDS
ONCODE INST HUBRECHT INST ROYAL NETHERLANDS ACAD ARTS & SCIENCES KNAW & UNIV MEDICAL CTR UTRECHT

RIVISTA

Biology Open, 8(8), bio042812

IMPACT FACTOR

1,980

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