Amy C Richards Grayson1, Anne M Doody, David Putnam. 1. Department of Biomedical Engineering, The School of Chemical and Biomolecular Engineering, 270 Olin Hall, Cornell University, Ithaca, New York 14853, USA.
Abstract
PURPOSE: The goals of this study were as follows: 1) to evaluate the efficacy of different polyethylenimine (PEI) structures for siRNA delivery in a model system, and 2) to determine the biophysical and structural characteristics of PEI that relate to siRNA delivery. MATERIALS AND METHODS: Biophysical characterization (effective diameter and zeta potential), cytotoxicities, relative binding affinities and in vitro transfection efficiencies were determined using nano-complexes formed from PEI's of 800, 25,000, (both branched) and 22,000 (linear) molecular weights at varying N:P ratios and siRNA concentrations. The HR5-CL11 cell line stably expressing luciferase was used as a model system in vitro. RESULTS: Successful siRNA delivery was observed within a very narrow window of conditions, and only with the 25,000 branched PEI at an N:P ratio of 6:1 and 8:1 and with 200 nM siRNA. While the zeta potential and size of PEI:siRNA complexes correlated to transfection efficacy in some cases, complex stability may also affect transfection efficacy. CONCLUSIONS: The ability of PEI to transfer functionally active siRNA to cells in culture is surprisingly dependent on its biophysical and structural characteristics when compared to its relative success and ease of use for DNA delivery.
PURPOSE: The goals of this study were as follows: 1) to evaluate the efficacy of different polyethylenimine (PEI) structures for siRNA delivery in a model system, and 2) to determine the biophysical and structural characteristics of PEI that relate to siRNA delivery. MATERIALS AND METHODS: Biophysical characterization (effective diameter and zeta potential), cytotoxicities, relative binding affinities and in vitro transfection efficiencies were determined using nano-complexes formed from PEI's of 800, 25,000, (both branched) and 22,000 (linear) molecular weights at varying N:P ratios and siRNA concentrations. The HR5-CL11 cell line stably expressing luciferase was used as a model system in vitro. RESULTS: Successful siRNA delivery was observed within a very narrow window of conditions, and only with the 25,000 branched PEI at an N:P ratio of 6:1 and 8:1 and with 200 nM siRNA. While the zeta potential and size of PEI:siRNA complexes correlated to transfection efficacy in some cases, complex stability may also affect transfection efficacy. CONCLUSIONS: The ability of PEI to transfer functionally active siRNA to cells in culture is surprisingly dependent on its biophysical and structural characteristics when compared to its relative success and ease of use for DNA delivery.
Authors: Olivia M Merkel; Meredith A Mintzer; Damiano Librizzi; Olga Samsonova; Tanja Dicke; Brian Sproat; Holger Garn; Peter J Barth; Eric E Simanek; Thomas Kissel Journal: Mol Pharm Date: 2010-08-02 Impact factor: 4.939
Authors: Paschalia M Mountziaris; David C Sing; Sue Anne Chew; Stephanie N Tzouanas; E Dennis Lehman; F Kurtis Kasper; Antonios G Mikos Journal: Pharm Res Date: 2010-12-24 Impact factor: 4.200
Authors: James E Dahlman; Carmen Barnes; Omar Khan; Aude Thiriot; Siddharth Jhunjunwala; Taylor E Shaw; Yiping Xing; Hendrik B Sager; Gaurav Sahay; Lauren Speciner; Andrew Bader; Roman L Bogorad; Hao Yin; Tim Racie; Yizhou Dong; Shan Jiang; Danielle Seedorf; Apeksha Dave; Kamaljeet S Sandu; Matthew J Webber; Tatiana Novobrantseva; Vera M Ruda; Abigail K R Lytton-Jean; Christopher G Levins; Brian Kalish; Dayna K Mudge; Mario Perez; Ludmila Abezgauz; Partha Dutta; Lynelle Smith; Klaus Charisse; Mark W Kieran; Kevin Fitzgerald; Matthias Nahrendorf; Dganit Danino; Rubin M Tuder; Ulrich H von Andrian; Akin Akinc; Avi Schroeder; Dipak Panigrahy; Victor Kotelianski; Robert Langer; Daniel G Anderson Journal: Nat Nanotechnol Date: 2014-05-11 Impact factor: 39.213