PMID- 38526634
OWN - NLM
STAT- Publisher
LR  - 20240325
IS  - 2190-3948 (Electronic)
IS  - 2190-393X (Linking)
DP  - 2024 Mar 25
TI  - The intracellular visualization of exogenous DNA in fluorescence microscopy.
LID - 10.1007/s13346-024-01563-4 [doi]
AB  - In the development of non-viral gene delivery vectors, it is essential to 
      reliably localize and quantify transfected DNA inside the cell. To track DNA, 
      fluorescence microscopy methods are commonly applied. These mostly rely on 
      fluorescently labeled DNA, DNA binding proteins fused to a fluorescent protein, 
      or fluorescence in situ hybridization (FISH). In addition, co-stainings are often 
      used to determine the colocalization of the DNA in specific cellular 
      compartments, such as the endolysosomes or the nucleus. We provide an overview of 
      these DNA tracking methods, advice on how they should be combined, and indicate 
      which co-stainings or additional methods are required to draw precise conclusions 
      from a DNA tracking experiment. Some emphasis is given to the localization of 
      exogenous DNA inside the nucleus, which is the last step of DNA delivery. We 
      argue that suitable tools which allow for the nuclear detection of faint signals 
      are still missing, hampering the rational development of more efficient non-viral 
      transfection systems.
CI  - (c) 2024. The Author(s).
FAU - Greitens, Christina
AU  - Greitens C
AUID- ORCID: 0009-0001-2231-3126
AD  - Institute of Pharmaceutical Sciences, Department of Chemistry and Applied 
      Biosciences, ETH Zurich, 8093, Zurich, Switzerland.
FAU - Leroux, Jean-Christophe
AU  - Leroux JC
AUID- ORCID: 0000-0001-5601-1292
AD  - Institute of Pharmaceutical Sciences, Department of Chemistry and Applied 
      Biosciences, ETH Zurich, 8093, Zurich, Switzerland. jleroux@ethz.ch.
FAU - Burger, Michael
AU  - Burger M
AUID- ORCID: 0000-0001-9253-0575
AD  - Institute of Pharmaceutical Sciences, Department of Chemistry and Applied 
      Biosciences, ETH Zurich, 8093, Zurich, Switzerland. 
      Michael.Burger@pharma.ethz.ch.
LA  - eng
GR  - 884505/HORIZON EUROPE European Research Council/
PT  - Journal Article
DEP - 20240325
PL  - United States
TA  - Drug Deliv Transl Res
JT  - Drug delivery and translational research
JID - 101540061
SB  - IM
OTO - NOTNLM
OT  - DNA visualization
OT  - DNA-binding proteins
OT  - Fluorescence in situ hybridization
OT  - Fluorescence microscopy
OT  - Gene delivery
OT  - Labeled DNA
OT  - Non-viral vectors
EDAT- 2024/03/25 18:42
MHDA- 2024/03/25 18:42
CRDT- 2024/03/25 12:06
PHST- 2024/02/28 00:00 [accepted]
PHST- 2024/03/25 18:42 [medline]
PHST- 2024/03/25 18:42 [pubmed]
PHST- 2024/03/25 12:06 [entrez]
AID - 10.1007/s13346-024-01563-4 [pii]
AID - 10.1007/s13346-024-01563-4 [doi]
PST - aheadofprint
SO  - Drug Deliv Transl Res. 2024 Mar 25. doi: 10.1007/s13346-024-01563-4.