Microarray Analysis of Gene Expression Profiles in Cells Transfected With Nonviral Vectors

Inefficient gene delivery is a critical factor limiting the use of nonviral methods in therapeutic applications including gene therapy and tissue engineering. There have been few efforts to understand or engineer the molecular signaling pathways that dictate the efficacy of gene transfer. Microarray...

Full description

Saved in:
Bibliographic Details
Published in:Molecular therapy 2011-12, Vol.19 (12), p.2144-2151
Main Authors: Plautz, Sarah A, Boanca, Gina, Riethoven, Jean-Jack M, Pannier, Angela K
Format: Article
Language:English
Subjects:
Animals
Biomarkers - metabolism
Cell adhesion & migration
Cells, Cultured
Cytotoxicity
Efficiency
Flow cytometry
Gene expression
Gene Expression Profiling
Gene therapy
Genetic Vectors - administration & dosage
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Humans
Lipids
Mice
NIH 3T3 Cells
Oligonucleotide Array Sequence Analysis
Original
Proteins
Real-Time Polymerase Chain Reaction
RNA, Messenger - genetics
Systems engineering
Tissue engineering
Transfection
Vectors (Biology)
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Inefficient gene delivery is a critical factor limiting the use of nonviral methods in therapeutic applications including gene therapy and tissue engineering. There have been few efforts to understand or engineer the molecular signaling pathways that dictate the efficacy of gene transfer. Microarray analysis was used to determine endogenous gene expression profiles modulated during nonviral gene transfer. Nonviral DNA lipoplexes were delivered to HEK 293T cells. Flow cytometry was used to isolate a population of transfected cells. Expression patterns were compared between transfected and nontransfected samples, which revealed three genes that were significantly upregulated in transfected cells, including RAP1A, a GTPase implicated in integrin-mediated cell adhesion, and HSP70B′, a stress-inducible gene that may be important for maintaining cell viability. Furthermore, RAP1A was also significantly upregulated in untransfected cells that were exposed to lipoplexes but that had not expressed the transgene as compared to control, untreated cells. Transfection in the presence of activators of upregulated genes was enhanced, demonstrating the principle of altering endogenous gene expression profiles to enhance transfection. With a greater understanding of signaling pathways involved in gene delivery, more efficient nonviral delivery schemes capitalizing on endogenous factors can be developed to advance therapeutic applications.
ISSN:1525-0016
1525-0024
DOI:10.1038/mt.2011.161