PMID- 34424689
OWN - NLM
STAT- MEDLINE
DCOM- 20220202
LR  - 20220202
IS  - 1520-5126 (Electronic)
IS  - 0002-7863 (Linking)
VI  - 143
IP  - 34
DP  - 2021 Sep 1
TI  - A Universal Labeling Strategy for Nucleic Acids in Expansion Microscopy.
PG  - 13782-13789
LID - 10.1021/jacs.1c05931 [doi]
AB  - Expansion microscopy (ExM) enables the nanoscale imaging of ribonucleic acids 
      (RNAs) on a conventional fluorescence microscope, providing information on the 
      intricate patterns of gene expression at (sub)cellular resolution and within 
      spatial context. To extend the use of such strategies, we examined a series of 
      multivalent reagents that allow the labeling and grafting of deoxyribonucleic 
      acid (DNA) oligonucleotide probes in a unified approach. We show that the 
      reagents are directly compatible with third-generation in situ hybridization 
      chain reaction RNA FISH (fluorescence in situ hybridization) techniques while 
      displaying complete retention of the targeted transcripts. Furthermore, we 
      validate and demonstrate that our labeling method is compatible with multicolor 
      staining. Through oligonucleotide-conjugated antibodies, we demonstrate excellent 
      performance in x4 ExM and x10 ExM, achieving a resolution of  approximately 50 nm in x10 ExM 
      for both pre- and postexpansion labeling strategies. Our results indicate that 
      our multivalent molecules enable the rapid functionalization of DNA 
      oligonucleotides for ExM.
FAU - Wen, Gang
AU  - Wen G
AD  - Department of Chemistry, KU Leuven, Leuven 3001, Belgium.
FAU - Vanheusden, Marisa
AU  - Vanheusden M
AD  - Department of Chemistry, KU Leuven, Leuven 3001, Belgium.
FAU - Leen, Volker
AU  - Leen V
AD  - Chrometra, Kortenaken, 3470 Belgium.
FAU - Rohand, Taoufik
AU  - Rohand T
AD  - Faculty Polydisciplinaire of Safi, Department of Chemistry, Laboratory of 
      Analytical & Molecular Chemistry, University Cadi Ayyad, 46000 Safi, Morocco.
FAU - Vandereyken, Katy
AU  - Vandereyken K
AD  - Department of Human Genetics, KU Leuven, Leuven 3000, Belgium.
AD  - LISCO, KU Leuven Institute for Single-Cell Omics, Leuven 3000, Belgium.
FAU - Voet, Thierry
AU  - Voet T
AD  - Department of Human Genetics, KU Leuven, Leuven 3000, Belgium.
AD  - LISCO, KU Leuven Institute for Single-Cell Omics, Leuven 3000, Belgium.
FAU - Hofkens, Johan
AU  - Hofkens J
AUID- ORCID: 0000-0002-9101-0567
AD  - Department of Chemistry, KU Leuven, Leuven 3001, Belgium.
AD  - LISCO, KU Leuven Institute for Single-Cell Omics, Leuven 3000, Belgium.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210823
PL  - United States
TA  - J Am Chem Soc
JT  - Journal of the American Chemical Society
JID - 7503056
RN  - 0 (Antibodies)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Nucleic Acids)
RN  - 0 (Oligonucleotides)
RN  - EC 2.7.2.3 (PGK1 protein, human)
RN  - EC 2.7.2.3 (Phosphoglycerate Kinase)
SB  - IM
MH  - Antibodies/chemistry
MH  - Fluorescent Dyes/chemistry
MH  - Gene Expression
MH  - HeLa Cells
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Microscopy, Fluorescence
MH  - Nucleic Acids/*chemistry
MH  - Oligonucleotides/chemistry
MH  - Phosphoglycerate Kinase/genetics/metabolism
MH  - Staining and Labeling/*methods
EDAT- 2021/08/24 06:00
MHDA- 2022/02/03 06:00
CRDT- 2021/08/23 17:26
PHST- 2021/08/24 06:00 [pubmed]
PHST- 2022/02/03 06:00 [medline]
PHST- 2021/08/23 17:26 [entrez]
AID - 10.1021/jacs.1c05931 [doi]
PST - ppublish
SO  - J Am Chem Soc. 2021 Sep 1;143(34):13782-13789. doi: 10.1021/jacs.1c05931. Epub 
      2021 Aug 23.