Journal article
Correlating intracellular nonviral polyplex localization with transfection efficiency using high-content screening
Biotechnology progress, Vol.31(6), pp.1685-1692
11/2015
DOI: 10.1002/btpr.2146
PMCID: PMC4783161
PMID: 26193826
Abstract
High-content screening (HCS) has gained interest in cellular imaging because of its ability to provide statistically significant data from multiple parameters simultaneously in cell-based assays. Although HCS has been mainly used in drug discovery, it has other potentially useful applications, such as elucidating the processes involved in nonviral gene vector-mediated gene delivery, as was explored in this study. HCS was used to measure transfection efficiency and cytotoxicities of polyplexes made from fluorescently labeled polyethylenimine (PEI) and pDNA encoding EGFP (pEGFP-N1). The results generated using HCS were confirmed using more conventional and labor-intensive methods. For the first time, a relationship between transfected cells and the number of polyplexes in the cytoplasm was shown. Four to five polyplex signals were found in the cytoplasm of successfully transfected cells, whilst nontransfected cells harbored less than one polyplex signal within the cytoplasm. HCS has the potential to be used as a tool in the field of gene delivery. HCS can not only simultaneously measure transfection efficiency and cytotoxicity of various nonviral gene vectors; it can also be used to track such vectors through various subcellular compartments.
Details
- Title: Subtitle
- Correlating intracellular nonviral polyplex localization with transfection efficiency using high-content screening
- Creators
- Amaraporn Wongrakpanich - Dept. of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, University of Iowa, Iowa City, IA, 52241Meng Wu - The University of Iowa, High Throughput Screening Facility (UIHTS), College of Pharmacy, University of Iowa, Iowa City, IA, 52241Aliasger K Salem - Dept. of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, University of Iowa, Iowa City, IA, 52241
- Resource Type
- Journal article
- Publication Details
- Biotechnology progress, Vol.31(6), pp.1685-1692
- DOI
- 10.1002/btpr.2146
- PMID
- 26193826
- PMCID
- PMC4783161
- NLM abbreviation
- Biotechnol Prog
- ISSN
- 8756-7938
- eISSN
- 1520-6033
- Publisher
- United States
- Grant note
- P30 ES005605 / NIEHS NIH HHS P50 CA97274 / NCI NIH HHS P30 CA086862 / NCI NIH HHS P50 CA097274 / NCI NIH HHS
- Language
- English
- Date published
- 11/2015
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Pharmacy; Core Research Facilities; Pharmaceutical Sciences and Experimental Therapeutics; Craniofacial Anomalies Research Center; Biochemistry and Molecular Biology; Dental Research; Chemical and Biochemical Engineering
- Record Identifier
- 9983985841902771
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