Electroporation and Microinjection Successfully Deliver Single-Stranded and Duplex DNA into Live Cells as Detected by FRET Measurements

Förster resonance energy transfer (FRET) technology relies on the close proximity of two compatible fluorophores for energy transfer. Tagged (Cy3 and Cy5) complementary DNA strands forming a stable duplex and a doubly-tagged single strand were shown to demonstrate FRET outside of a cellular environm...

Descripción completa

Detalles Bibliográficos
Autores principales: Bamford, Rosemary A., Zhao, Zheng-yun, Hotchin, Neil A., Styles, Iain B., Nash, Gerard B., Tucker, James H. R., Bicknell, Roy
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3995676/
https://www.ncbi.nlm.nih.gov/pubmed/24755680
http://dx.doi.org/10.1371/journal.pone.0095097
_version_ 1782312913684398080
author Bamford, Rosemary A.
Zhao, Zheng-yun
Hotchin, Neil A.
Styles, Iain B.
Nash, Gerard B.
Tucker, James H. R.
Bicknell, Roy
author_facet Bamford, Rosemary A.
Zhao, Zheng-yun
Hotchin, Neil A.
Styles, Iain B.
Nash, Gerard B.
Tucker, James H. R.
Bicknell, Roy
author_sort Bamford, Rosemary A.
collection PubMed
description Förster resonance energy transfer (FRET) technology relies on the close proximity of two compatible fluorophores for energy transfer. Tagged (Cy3 and Cy5) complementary DNA strands forming a stable duplex and a doubly-tagged single strand were shown to demonstrate FRET outside of a cellular environment. FRET was also observed after transfecting these DNA strands into fixed and live cells using methods such as microinjection and electroporation, but not when using lipid based transfection reagents, unless in the presence of the endosomal acidification inhibitor bafilomycin. Avoiding the endocytosis pathway is essential for efficient delivery of intact DNA probes into cells.
format Online
Article
Text
id pubmed-3995676
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-39956762014-04-25 Electroporation and Microinjection Successfully Deliver Single-Stranded and Duplex DNA into Live Cells as Detected by FRET Measurements Bamford, Rosemary A. Zhao, Zheng-yun Hotchin, Neil A. Styles, Iain B. Nash, Gerard B. Tucker, James H. R. Bicknell, Roy PLoS One Research Article Förster resonance energy transfer (FRET) technology relies on the close proximity of two compatible fluorophores for energy transfer. Tagged (Cy3 and Cy5) complementary DNA strands forming a stable duplex and a doubly-tagged single strand were shown to demonstrate FRET outside of a cellular environment. FRET was also observed after transfecting these DNA strands into fixed and live cells using methods such as microinjection and electroporation, but not when using lipid based transfection reagents, unless in the presence of the endosomal acidification inhibitor bafilomycin. Avoiding the endocytosis pathway is essential for efficient delivery of intact DNA probes into cells. Public Library of Science 2014-04-22 /pmc/articles/PMC3995676/ /pubmed/24755680 http://dx.doi.org/10.1371/journal.pone.0095097 Text en © 2014 Bamford et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Bamford, Rosemary A.
Zhao, Zheng-yun
Hotchin, Neil A.
Styles, Iain B.
Nash, Gerard B.
Tucker, James H. R.
Bicknell, Roy
Electroporation and Microinjection Successfully Deliver Single-Stranded and Duplex DNA into Live Cells as Detected by FRET Measurements
title Electroporation and Microinjection Successfully Deliver Single-Stranded and Duplex DNA into Live Cells as Detected by FRET Measurements
title_full Electroporation and Microinjection Successfully Deliver Single-Stranded and Duplex DNA into Live Cells as Detected by FRET Measurements
title_fullStr Electroporation and Microinjection Successfully Deliver Single-Stranded and Duplex DNA into Live Cells as Detected by FRET Measurements
title_full_unstemmed Electroporation and Microinjection Successfully Deliver Single-Stranded and Duplex DNA into Live Cells as Detected by FRET Measurements
title_short Electroporation and Microinjection Successfully Deliver Single-Stranded and Duplex DNA into Live Cells as Detected by FRET Measurements
title_sort electroporation and microinjection successfully deliver single-stranded and duplex dna into live cells as detected by fret measurements
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3995676/
https://www.ncbi.nlm.nih.gov/pubmed/24755680
http://dx.doi.org/10.1371/journal.pone.0095097
work_keys_str_mv AT bamfordrosemarya electroporationandmicroinjectionsuccessfullydeliversinglestrandedandduplexdnaintolivecellsasdetectedbyfretmeasurements
AT zhaozhengyun electroporationandmicroinjectionsuccessfullydeliversinglestrandedandduplexdnaintolivecellsasdetectedbyfretmeasurements
AT hotchinneila electroporationandmicroinjectionsuccessfullydeliversinglestrandedandduplexdnaintolivecellsasdetectedbyfretmeasurements
AT stylesiainb electroporationandmicroinjectionsuccessfullydeliversinglestrandedandduplexdnaintolivecellsasdetectedbyfretmeasurements
AT nashgerardb electroporationandmicroinjectionsuccessfullydeliversinglestrandedandduplexdnaintolivecellsasdetectedbyfretmeasurements
AT tuckerjameshr electroporationandmicroinjectionsuccessfullydeliversinglestrandedandduplexdnaintolivecellsasdetectedbyfretmeasurements
AT bicknellroy electroporationandmicroinjectionsuccessfullydeliversinglestrandedandduplexdnaintolivecellsasdetectedbyfretmeasurements

Ejemplares similares