Evidence map›Paper›PMID 25753113›Full record

ArticleBiochimica et biophysica acta2015

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

Ramsey N Majzoub, Chia-Ling Chan, Kai K Ewert, Bruno F B Silva, Keng S Liang, Cyrus R Safinya

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Biochimica et biophysica acta, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.

0numbers the graph read from it
0cells of the map it votes in
17citing papers in PubMed
1.7field-weighted citation impact, top 16% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

17 citing papers in PubMed, 31 citations in OpenAlex.

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  12. Cationic liposome-nucleic acid nanoparticle assemblies with applications in gene delivery and gene silencing.Philosophical transactions. Series A, Mathematical, physical, and engineering sciences · 2016
    Review
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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors at 2 institutions in 3 countries.

Ramsey N MajzoubDepartment of Physics, University of California, Santa Barbara, CA 93106, USA; Department of Materials, University of California, Santa Barbara, CA 93106, USA; Molecular, Cellular and Developmental Biology Department, University of California, Santa Barbara, CA 93106, USA.
Chia-Ling ChanDepartment of Physics, University of California, Santa Barbara, CA 93106, USA; Department of Materials, University of California, Santa Barbara, CA 93106, USA; Molecular, Cellular and Developmental Biology Department, University of California, Santa Barbara, CA 93106, USA; Institute of Physics, Academica Sinica, Taipei 11529, Taiwan; National Synchrotron Radiation Research Center, Hsinchu 30076, Taiwan.
Kai K EwertDepartment of Physics, University of California, Santa Barbara, CA 93106, USA; Department of Materials, University of California, Santa Barbara, CA 93106, USA; Molecular, Cellular and Developmental Biology Department, University of California, Santa Barbara, CA 93106, USA.
Bruno F B SilvaDepartment of Physics, University of California, Santa Barbara, CA 93106, USA; Department of Materials, University of California, Santa Barbara, CA 93106, USA; Molecular, Cellular and Developmental Biology Department, University of California, Santa Barbara, CA 93106, USA; Division of Physical Chemistry, Centre for Chemistry and Chemical Engineering, Lund University, SE-221 00 Lund, Sweden.
Keng S LiangNational Synchrotron Radiation Research Center, Hsinchu 30076, Taiwan; Department of Electrophysics, National Chiao-Tung University, Hsinchu 30010, Taiwan.
Cyrus R SafinyaDepartment of Physics, University of California, Santa Barbara, CA 93106, USA; Department of Materials, University of California, Santa Barbara, CA 93106, USA; Molecular, Cellular and Developmental Biology Department, University of California, Santa Barbara, CA 93106, USA. Electronic address: safinya@mrl.ucsb.edu.
University of California, Santa Barbara · USNational Yang Ming Chiao Tung University · TW

Funding

Structures of Lipid-DNA Nonviral Gene Delivery Systems 4R01GM059288 · NIGMS · UNIVERSITY OF CALIFORNIA SANTA BARBARA · PI SAFINYA, CYRUS R · 1999 to 2016
$4.2M
NIGMS NIH HHS GM-59288NIGMS NIH HHS R01 GM059288
6 · The paper itself

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 Rab5-GDP to the plasma membrane and not indicative of NP escape from EEs. Taken together, our results show that endosomal escape of PEGylated nanoparticles occurs downstream of EEs i.e., from late endosomes/lysosomes. Our studies also suggest that Rab5-Q79L could be used in a robust imaging assay which allows for direct visualization of NP interactions with the luminal membrane of early endosomes.

Indexed as

AnimalsCationsCell LineEndosomesGreen Fluorescent ProteinsLipidsLiposomesMiceMicroscopy, FluorescenceModels, BiologicalMutant ProteinsNanoparticlesNucleic AcidsParticle SizePolyethylene Glycolsrab5 GTP-Binding ProteinsCationsGreen Fluorescent ProteinsLipidsLiposomesMutant ProteinsNucleic AcidsPolyethylene Glycolsrab5 GTP-Binding ProteinsRecombinant Fusion ProteinsEarly endosomesFluorescence imagingGene deliveryLipid–DNA nanoparticlesParticle colocalizationRab GTPases

Identifiers

PMID25753113
PMCPMC4398653
OpenAlexW2157193154

What OpenQuestion holds

Textmetadata
LicenceTDM
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.