PMID- 37753471
OWN - NLM
STAT- Publisher
LR  - 20230928
IS  - 2331-8325 (Electronic)
IS  - 2331-8325 (Linking)
VI  - 13
IP  - 18
DP  - 2023 Sep 20
TI  - Fluorescence in situ Localization of Pri-miRNAs in Isolated Arabidopsis thaliana 
      Nuclei.
PG  - e4824
LID - 10.21769/BioProtoc.4824 [doi]
LID - e4824
AB  - Here, we present an approach combining fluorescence in situ hybridization (FISH) 
      and immunolabeling for localization of pri-miRNAs in isolated nuclei of A. 
      thaliana. The presented method utilizes specific DNA oligonucleotide probes, 
      modified by addition of digoxigenin-labeled deoxynucleotides to its 3' hydroxyl 
      terminus by terminal deoxynucleotidyl transferase (TdT). The probes are then 
      detected by immunolabeling of digoxigenin (DIG) using specific 
      fluorescent-labeled antibodies to visualize hybridized probes. Recently, we have 
      applied this method to localize pri-miRNA156a, pri-miRNA163, pri-miRNA393a, and 
      pri-miRNA414 in the nuclei isolated from leaves of 4-week-old A. thaliana. The 
      present approach can be easily implemented to analyze nuclear distribution of 
      diverse RNA classes, including mRNAs and pri-miRNAs in isolated fixed cells or 
      nuclei from plant.
CI  - (c)Copyright : (c) 2023 The Authors; This is an open access article under the CC 
      BY-NC license.
FAU - Gulanicz, Tomasz
AU  - Gulanicz T
AD  - Centre for Modern Interdisciplinary Technologies, Nicolaus Copernicus University, 
      Torun, Poland.
FAU - Zienkiewicz, Agnieszka
AU  - Zienkiewicz A
AD  - Centre for Modern Interdisciplinary Technologies, Nicolaus Copernicus University, 
      Torun, Poland.
FAU - Zienkiewicz, Krzysztof
AU  - Zienkiewicz K
AD  - Centre for Modern Interdisciplinary Technologies, Nicolaus Copernicus University, 
      Torun, Poland.
FAU - Kasprowicz-Maluski, Anna
AU  - Kasprowicz-Maluski A
AD  - Department of Molecular and Cellular Biology, Institute of Molecular Biology and 
      Biotechnology, Adam Mickiewicz University, Poznan, Poland.
FAU - Szweykowska-Kulinska, Zofia
AU  - Szweykowska-Kulinska Z
AD  - Department of Gene Expression, Institute of Molecular Biology and Biotechnology, 
      Adam Mickiewicz University, Poznan, Poland.
FAU - Jarmolowski, Artur
AU  - Jarmolowski A
AD  - Department of Gene Expression, Institute of Molecular Biology and Biotechnology, 
      Adam Mickiewicz University, Poznan, Poland.
LA  - eng
PT  - Journal Article
DEP - 20230920
PL  - United States
TA  - Bio Protoc
JT  - Bio-protocol
JID - 101635102
PMC - PMC10518776
OTO - NOTNLM
OT  - Confocal microscopy
OT  - Digoxigenin
OT  - Fluorescent in situ hybridization
OT  - Immunolabeling
OT  - Pri-miRNA
COIS- Competing interestsThere are no conflicts of interest or competing interests.
EDAT- 2023/09/27 06:42
MHDA- 2023/09/27 06:42
PMCR- 2023/09/20
CRDT- 2023/09/27 03:51
PHST- 2023/03/20 00:00 [received]
PHST- 2023/06/25 00:00 [revised]
PHST- 2023/07/17 00:00 [accepted]
PHST- 2023/09/27 06:42 [medline]
PHST- 2023/09/27 06:42 [pubmed]
PHST- 2023/09/27 03:51 [entrez]
PHST- 2023/09/20 00:00 [pmc-release]
AID - e4824 [pii]
AID - 4824 [pii]
AID - 10.21769/BioProtoc.4824 [doi]
PST - epublish
SO  - Bio Protoc. 2023 Sep 20;13(18):e4824. doi: 10.21769/BioProtoc.4824. eCollection 
      2023 Sep 20.