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Enrichment strategies for siRNA-transfected cells in an untransfected background

Paper ID Volume ID Publish Year Pages File Format Full-Text
25214 43561 2007 4 PDF Available
Title
Enrichment strategies for siRNA-transfected cells in an untransfected background
Abstract

Gene silencing experiments in difficult-to-transfect cells are often hampered by the presence of a background of untransfected cells. We present proof-of-concept data from two different strategies for enrichment of siRNA-transfected cells. In the first approach, a heterologous surface antigen is expressed from a plasmid that is co-transfected with an siRNA targeting an endogenous mRNA. The surface antigen is then used for enrichment of successfully transfected cells using antibody-coated magnetic particles. In the second strategy, a eukaryotic antibiotic resistance gene is expressed from a co-transfected plasmid. Addition of the corresponding antibiotic 24 h after transfection results in killing of untransfected cells, which can be washed away. Elimination of untransfected cells will allow more accurate interpretation of the effects of gene silencing.

Keywords
GAPDH, glyceraldehyde-3-phosphate dehydrogenase; MAPK1, mitogen-activated protein kinase 1siRNA; Transfection; Cell separation
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Enrichment strategies for siRNA-transfected cells in an untransfected background
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 130, Issue 3, 30 June 2007, Pages 209–212
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