Fluorescence microscopy colocalization of lipid-nucleic acid nanoparticles with wildtype and mutant Rab5-GFP: A platform for investigating early endosomal events

Biochimica Et Biophysica Acta
Ramsey N MajzoubCyrus R Safinya

Abstract

Endosomal entrapment is known to be a major bottleneck to successful cytoplasmic delivery of nucleic acids (NAs) using cationic liposome-NA nanoparticles (NPs). Quantitative measurements of distributions of NPs within early endosomes (EEs) have proven difficult due to the sub-resolution size and short lifetime of wildtype EEs. In this study we used Rab5-GFP, a member of the large family of GTPases which cycles between the plasma membrane and early endosomes, to fluorescently label early endosomes. Using fluorescence microscopy and quantitative image analysis of cells expressing Rab5-GFP, we found that at early time points (t<1h), only a fraction (≈35%) of RGD-tagged NPs (which target cell surface integrins) colocalize with wildtype EEs, independent of the NP's membrane charge density. In comparison, a GTP-hydrolysis deficient mutant, Rab5-Q79L, which extends the size and lifetime of EEs yielding giant early endosomes (GEEs), enabled us to resolve and localize individual NPs found within the GEE lumen. Remarkably, nearly all intracellular NPs are found to be trapped within GEEs implying little or no escape at early time points. The observed small degree of colocalization of NPs and wildtype Rab5 is consistent with recycling of R...Continue Reading

References

Mar 8, 1991·Cell·J P GorvelJ Gruenberg
Jan 1, 1996·Annual Review of Cell and Developmental Biology·E Ruoslahti
Nov 17, 1999·Proceedings of the Society for Experimental Biology and Medicine·G J Mizejewski
Jul 12, 2003·Journal of Liposome Research·Yasuyuki SadzukaTakashi Sonobe
Jan 16, 2004·Trends in Pharmacological Sciences·Mouldy Sioud
Jan 11, 2005·Current Gene Therapy·Youngsuk YiKwan Hee Lee
Feb 3, 2005·Nature Reviews. Drug Discovery·Vladimir P Torchilin
Feb 16, 2005·Expert Opinion on Biological Therapy·Kai K EwertCyrus R Safinya
Jun 1, 2005·Chemistry and Physics of Lipids·Olga GarbuzenkoAba Priev
Dec 21, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Susumu HamaHideyoshi Harashima
Apr 11, 2006·Journal of Immunological Methods·Jiyi YinTerri H Finkel
Jun 20, 2006·Biochimica Et Biophysica Acta·Akitada IwasaHideyoshi Harashima
Sep 6, 2006·Journal of Liposome Research·Joanna RejmanDick Hoekstra
Feb 13, 2007·Nanomedicine : Nanotechnology, Biology, and Medicine·Otilia M KooHayat Onyuksel
Jun 14, 2008·The Journal of Biological Chemistry·Ayman El-SayedHideyoshi Harashima
Sep 19, 2008·Colloids and Surfaces. B, Biointerfaces·Minh-Uyen TrinhDaniel Fornasiero
Jul 16, 2009·Nature Reviews. Molecular Cell Biology·Harald Stenmark
Dec 8, 2009·Cancer Cell·Kazuki N SugaharaErkki Ruoslahti
Mar 17, 2010·The Journal of Cell Biology·Erkki RuoslahtiMichael J Sailor
Jul 27, 2010·Journal of Drug Targeting·Federico PerchePatrick Midoux
Oct 20, 2010·International Journal of Nanomedicine·Torben GjettingHans Skovgaard Poulsen
Jul 27, 2011·Journal of Controlled Release : Official Journal of the Controlled Release Society·Zia ur RehmanInge S Zuhorn
Aug 30, 2011·Accounts of Chemical Research·Xia Guo, Leaf Huang

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Citations

Jun 7, 2015·Langmuir : the ACS Journal of Surfaces and Colloids·Ramsey N MajzoubCyrus R Safinya
Nov 7, 2015·Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology·Martin P StewartGaurav Sahay
Jun 15, 2016·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Ramsey N MajzoubCyrus R Safinya
Feb 15, 2016·Bioorganic & Medicinal Chemistry Letters·Kai K EwertCyrus R Safinya
Feb 2, 2018·Advanced Materials·Joël BourquinBarbara Rothen-Rutishauser
Nov 1, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Ricardo GasparBruno F B Silva
Jun 3, 2021·Journal of Controlled Release : Official Journal of the Controlled Release Society·Emily XuAlexandra S Piotrowski-Daspit
May 16, 2019·Chemical Reviews·Patrick G DoughertyDehua Pei
Jul 8, 2017·Biomacromolecules·Che Fu LiuJin Kim Montclare

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