PMID- 28404480
OWN - NLM
STAT- MEDLINE
DCOM- 20180309
LR  - 20180522
IS  - 1095-9130 (Electronic)
IS  - 1046-2023 (Linking)
VI  - 120
DP  - 2017 May 1
TI  - 2D and 3D FISH of expanded repeat RNAs in human lymphoblasts.
PG  - 49-57
LID - S1046-2023(16)30386-3 [pii]
LID - 10.1016/j.ymeth.2017.04.002 [doi]
AB  - The first methods for visualizing RNAs within cells were designed for simple 
      imaging of specific transcripts in cells or tissues and since then significant 
      technical advances have been made in this field. Today, high-resolution images 
      can be obtained, enabling visualization of single transcript molecules, 
      quantitative analyses of images, and precise localization of RNAs within cells as 
      well as co-localization of transcripts with specific proteins or other molecules. 
      In addition, tracking of RNA dynamics within single cell has become possible. RNA 
      imaging techniques have been utilized for investigating the role of mutant RNAs 
      in a number of human disorders caused by simple microsatellite expansions. These 
      diseases include myotonic dystrophy type 1 and 2, amyotrophic lateral 
      sclerosis/frontotemporal dementia, fragile X-associated tremor/ataxia syndrome, 
      and Huntington's disease. Mutant RNAs with expanded repeats tend to aggregate 
      predominantly within cell nuclei, forming structures called RNA foci. In this 
      study, we demonstrate methods for fluorescent visualization of RNAs in both fixed 
      and living cells using the example of RNAs containing various expanded repeat 
      tracts (CUG, CCUG, GGGGCC, CGG, and CAG) from experiment design to image 
      analysis. We describe in detail 2D and 3D fluorescence in situ hybridization 
      (FISH) protocols for imaging expanded repeats RNAs, and we review briefly live 
      imaging techniques used to characterize RNA foci formed by mutant RNAs. These 
      methods could be used to image the entire cellular pathway of RNAs, from 
      transcription to degradation.
CI  - Copyright (c) 2017 Elsevier Inc. All rights reserved.
FAU - Urbanek, Martyna O
AU  - Urbanek MO
AD  - Department of Molecular Biomedicine, Institute of Bioorganic Chemistry, Polish 
      Academy of Sciences, Noskowskiego 12/14 Str., 61-704 Poznan, Poland.
FAU - Michalak, Michal
AU  - Michalak M
AD  - Department of Molecular Biomedicine, Institute of Bioorganic Chemistry, Polish 
      Academy of Sciences, Noskowskiego 12/14 Str., 61-704 Poznan, Poland.
FAU - Krzyzosiak, Wlodzimierz J
AU  - Krzyzosiak WJ
AD  - Department of Molecular Biomedicine, Institute of Bioorganic Chemistry, Polish 
      Academy of Sciences, Noskowskiego 12/14 Str., 61-704 Poznan, Poland. Electronic 
      address: wlodkrzy@ibch.poznan.pl.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20170409
PL  - United States
TA  - Methods
JT  - Methods (San Diego, Calif.)
JID - 9426302
RN  - 0 (Fluorescent Dyes)
RN  - 63231-63-0 (RNA)
RN  - Fragile X Tremor Ataxia Syndrome
RN  - Frontotemporal Dementia With Motor Neuron Disease
SB  - IM
MH  - Amyotrophic Lateral Sclerosis/genetics
MH  - Ataxia/genetics
MH  - Cells, Cultured
MH  - Fluorescent Dyes/*chemistry
MH  - Fragile X Syndrome/genetics
MH  - Frontotemporal Dementia/genetics
MH  - Humans
MH  - Huntington Disease/genetics
MH  - In Situ Hybridization, Fluorescence/instrumentation/*methods
MH  - Lymphocytes
MH  - Microscopy, Confocal
MH  - Molecular Imaging/instrumentation/*methods
MH  - Myotonic Dystrophy/genetics
MH  - RNA/*chemistry/genetics
MH  - Tremor/genetics
MH  - *Trinucleotide Repeat Expansion
OTO - NOTNLM
OT  - Fluorescence in situ hybridization
OT  - RNA live imaging
OT  - RNA toxicity
OT  - Ribonuclear foci
OT  - Simple repeat expansion diseases
EDAT- 2017/04/14 06:00
MHDA- 2018/03/10 06:00
CRDT- 2017/04/14 06:00
PHST- 2016/12/15 00:00 [received]
PHST- 2017/02/09 00:00 [revised]
PHST- 2017/04/05 00:00 [accepted]
PHST- 2017/04/14 06:00 [pubmed]
PHST- 2018/03/10 06:00 [medline]
PHST- 2017/04/14 06:00 [entrez]
AID - S1046-2023(16)30386-3 [pii]
AID - 10.1016/j.ymeth.2017.04.002 [doi]
PST - ppublish
SO  - Methods. 2017 May 1;120:49-57. doi: 10.1016/j.ymeth.2017.04.002. Epub 2017 Apr 9.