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Multilayer mediated forward and patterned siRNA transfection using linear-PEI at extended N/P ratios

Paper ID Volume ID Publish Year Pages File Format Full-Text
1303 81 2009 15 PDF Available
Title
Multilayer mediated forward and patterned siRNA transfection using linear-PEI at extended N/P ratios
Abstract

Gene delivery from a substrate depends, in part, on the vector–nucleic acid complex that is bound to the surface and the cell adhesive properties of the surface. Here, we present a method to deliver patterns of small interfering RNA (siRNA) that capitalize on a forward transfection method (transfection by introducing siRNA transfection reagent complexes onto plated cells); herein denoted as multilayer mediated forward transfection (MFT). This method separates the substrate-mediated delivery from the cell adhesive properties of the surface. pH responsive layer-by-layer (LbL) assembled multilayers were used as the delivery platform and microcontact printing technique (μCP) was used to pattern nanoparticles of transfection reagent–siRNA complexes onto degradable multilayers. Efficient MFT depend on optimal formulation of the nanoparticles. 25 kDa linear polyethylenimine (LPEI) was optimized as the siRNA transfection reagent for normal forward transfection (NFT) of the nanoparticles. A broad range of LPEI–siRNA nitrogen/phosphate (N/P) ratios (ranging from 5 to 90) was evaluated for the relative amounts of siRNA incorporated into the nanoparticles, nanoparticle size and NFT efficiencies. All the siRNA was incorporated into the nanoparticles at N/P ratio near 90. Increasing the amount of siRNA incorporated into the nanoparticles, with increasing N/P ratio correlated with a linear blue shift in the ultraviolet/visible (UV/vis) absorbance spectrum of the LPEI–siRNA nanoparticles. NFT efficiency greater than 80% was achieved with minimal cytotoxicity at N/P ratio of 30 and siRNA concentration of 200 nM. Similarly, MFT efficiency ⩾60% was achieved for LPEI–siRNA nanoparticles at N/P ratios greater than 30.

Keywords
siRNA transfection; Patterned transfection; Layer-by-layer (LbL) assembly; LPEI–siRNA nanoparticles; N/P ratio
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Multilayer mediated forward and patterned siRNA transfection using linear-PEI at extended N/P ratios
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Publisher
Database: Elsevier - ScienceDirect
Journal: Acta Biomaterialia - Volume 5, Issue 5, June 2009, Pages 1474–1488
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us