Abstract:
:Here, we describe a robust protocol for the reverse transfection of cells on small interfering (siRNA) arrays, which, in combination with multi-channel immunofluorescence or time-lapse microscopy, is suitable for genome-wide RNA interference (RNAi) screens in intact human cells. The automatic production of 48 'transfection ready' siRNA arrays, each containing 384 samples, takes in total 7 h. Pre-fabricated siRNA arrays can be used without loss of transfection efficiency at least up to 15 months after printing. Different human cell lines that have been successfully transfected using the protocol are presented here. The present protocol has been applied to two genome-wide siRNA screens addressing mitosis and constitutive protein secretion.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Erfle H,Neumann B,Liebel U,Rogers P,Held M,Walter T,Ellenberg J,Pepperkok Rdoi
10.1038/nprot.2006.483subject
Has Abstractpub_date
2007-01-01 00:00:00pages
392-9issue
2eissn
1754-2189issn
1750-2799pii
nprot.2006.483journal_volume
2pub_type
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