Hi Dr. Ranjan,
Thank you so much for the informative and helpful response and for sharing the articles.
Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
Original Message:
Sent: 05-07-2025 16:20
From: Amalendu Ranjan
Subject: Question about siRNA-LNP endosomal escape
Hi Sheyda,
Yes, quantitatively measuring endosomal escape of siRNA-loaded lipid nanoparticles (siRNA-LNPs) is challenging. Using confocal microscopy is a great direct fluorescence method for quantification using co-localization of fluorescence using endosomal/lysosomal markers. The challenge is that colocalization is very low and it requires rigorous image analysis. Here are some reference articles relevant to this context
Lipid nanoparticle topology regulates endosomal escape and delivery of RNA to the cytoplasm | PNAS
The alternative approach may be gold siRNA tagged LNPs images by transmission electron microscopy.
Image-based analysis of lipid nanoparticle-mediated siRNA delivery, intracellular trafficking and endosomal escape - PubMed
Or Galectin-8 visualization for quantification as an alternative. Galectin 8 binds to exposed glycans on damaged endosomes. It is used as a marker for endosomal membrane rupture. In this article they have used for quantification
Gal8 Visualization of Endosome Disruption Predicts Carrier-Mediated Biologic Drug Intracellular Bioavailability
Usually combining two methods is ideal for validation.
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Amalendu Ranjan PhD
Assistant Professor
University of North Texas Health Science Center
Fort Worth TX
[email protected]
Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
Original Message:
Sent: 05-05-2025 12:16
From: Sheyda Ranjbar
Subject: Question about siRNA-LNP endosomal escape
Dear Nanotechnology community,
I'm looking for suggestions on how to quantitatively measure the endosomal escape of siRNA-loaded lipid nanoparticles (siRNA-LNPs). I've tried using confocal microscopy, but found it challenging to obtain reliable quantitative data. Any recommendations or alternative approaches would be greatly appreciated!
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Sheyda Ranjbar
Graduate Assistant
University of Connecticut School of Pharmacy
Storrs Mansfield CT
[email protected]
Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
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