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A simple and cost-effective method to transfect small interfering RNAs into pancreatic cancer cell lines using polyethylenimine.
MedLine Citation:
PMID:  20938367     Owner:  NLM     Status:  In-Process    
Abstract/OtherAbstract:
OBJECTIVES: RNA interference, an indispensable tool in functional genomics, can be induced by small interfering RNAs (siRNAs). Because of the transient nature of siRNA-mediated RNA interference, the continuous use of transfection reagents is mandatory. Because transfection reagents are expensive, cost-effective alternatives must be considered. In this study, we describe a polyethylenimine-based siRNA transfection protocol for pancreatic cancer cell lines.
METHODS: For determination of polyethylenimine-based transfection efficiency, a FAM-labeled siRNA was transfected into several pancreatic cancer cell lines and subsequently analyzed by flow cytometry. The effective knockdown of 2 siRNAs was determined on the protein level by Western blot. Toxicity of the transfection reagent was analyzed by viability assays.
RESULTS: Polyethylenimine can be used without overt cellular morphological changes, and toxicity is negligible in human and murine pancreatic cancer cell lines. Transfection efficiencies ranged between 83% and 98% in the cell lines used. The knockdown at the protein level was comparable to commercially available transfection reagents. Polyethylenimine and siRNA concentrations, incubation time, and cell density are determinates of the transfection efficiency.
CONCLUSIONS: Polyethylenimine is a suitable and cost-effective alternative for transfecting siRNAs into pancreatic cancer cells.
Authors:
Matthias Wirth; Petra Fritsche; Natasa Stojanovic; Martina Brandl; Stephanie Jaeckel; Roland M Schmid; Dieter Saur; Günter Schneider
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Pancreas     Volume:  40     ISSN:  1536-4828     ISO Abbreviation:  Pancreas     Publication Date:  2011 Jan 
Date Detail:
Created Date:  2010-12-16     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8608542     Medline TA:  Pancreas     Country:  United States    
Other Details:
Languages:  eng     Pagination:  144-50     Citation Subset:  IM    
Affiliation:
Technische Universität München, Klinikum rechts der Isar, II. Medizinische Klinik, München, Germany.
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