TY - JOUR
T1 - ECHO-liveFISH
T2 - In vivo RNA labeling reveals dynamic regulation of nuclear RNA foci in living tissues
AU - Oomoto, Ikumi
AU - Suzuki-Hirano, Asuka
AU - Umeshima, Hiroki
AU - Han, Yong Woon
AU - Yanagisawa, Hiroyuki
AU - Carlton, Peter
AU - Harada, Yoshie
AU - Kengaku, Mineko
AU - Okamoto, Akimitsu
AU - Shimogori, Tomomi
AU - Wang, Dan Ohtan
N1 - Publisher Copyright:
© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.
PY - 2015/10/30
Y1 - 2015/10/30
N2 - Elucidating the dynamic organization of nuclear RNA foci is important for understanding and manipulating these functional sites of gene expression in both physiological and pathological states. However, such studies have been difficult to establish in vivo as a result of the absence of suitable RNA imaging methods. Here, we describe a high-resolution fluorescence RNA imaging method, ECHO-liveFISH, to label endogenous nuclear RNA in living mice and chicks. Upon in vivo electroporation, exciton-controlled sequence-specific oligonucleotide probes revealed focally concentrated endogenous 28S rRNA and U3 snoRNA at nucleoli and poly(A) RNA at nuclear speckles. Time-lapse imaging reveals steady-state stability of these RNA foci and dynamic dissipation of 28S rRNA concentrations upon polymerase I inhibition in native brain tissue. Confirming the validity of this technique in a physiological context, the in vivo RNA labeling did not interfere with the function of target RNA nor cause noticeable cytotoxicity or perturbation of cellular behavior.
AB - Elucidating the dynamic organization of nuclear RNA foci is important for understanding and manipulating these functional sites of gene expression in both physiological and pathological states. However, such studies have been difficult to establish in vivo as a result of the absence of suitable RNA imaging methods. Here, we describe a high-resolution fluorescence RNA imaging method, ECHO-liveFISH, to label endogenous nuclear RNA in living mice and chicks. Upon in vivo electroporation, exciton-controlled sequence-specific oligonucleotide probes revealed focally concentrated endogenous 28S rRNA and U3 snoRNA at nucleoli and poly(A) RNA at nuclear speckles. Time-lapse imaging reveals steady-state stability of these RNA foci and dynamic dissipation of 28S rRNA concentrations upon polymerase I inhibition in native brain tissue. Confirming the validity of this technique in a physiological context, the in vivo RNA labeling did not interfere with the function of target RNA nor cause noticeable cytotoxicity or perturbation of cellular behavior.
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U2 - 10.1093/nar/gkv614
DO - 10.1093/nar/gkv614
M3 - Article
C2 - 26101260
AN - SCOPUS:84959329928
SN - 0305-1048
VL - 43
SP - e126
JO - Nucleic acids research
JF - Nucleic acids research
IS - 19
ER -