High-throughput RNA interference screens in Drosophila tissue culture cells.
This chapter describes the method used to conduct high-throughput screening (HTs) by RNA interference in Drosophila tissue culture cells. It covers four main topics: (1) a brief description of the existing platforms to conduct RNAi-screens in cell-based assays; (2) a table of the Drosophila cell lines available for these screens and a brief mention of the need to establish other cell lines as well as cultures of primary cells; (3) a discussion of the considerations and protocols involved in establishing assays suitable for HTS in a 384-well format; and (A) a summary of the various ways of handling raw data from an ongoing screen, with special emphasis on how to apply normalization for experimental variation and statistical filters to sort out noise from signals.
Armknecht
Susan
S
Department of Genetics, Harvard Medical School, Drosophila RNAi Screening Center, Boston, MA 02115, USA.
Boutros
Michael
M
Kiger
Amy
A
Nybakken
Kent
K
Mathey-Prevot
Bernard
B
Perrimon
Norbert
N
D000818:Animals; D002460:Cell Line; D004330:Drosophila / Q000166:cytology / Q000235:genetics*; D017930:Genes, Reporter; D034622:RNA Interference; D046509:Tissue Culture Techniques; D014162:Transfection