title: Transgene-induced RNA interference as a tool for plant functional genomics.
abstract: RNA interference (RNAi) is a powerful tool for functional genomics in a number of species. The logistics and procedures for doing high-throughput RNAi to investigate the functions of large numbers of genes in Arabidopsis thaliana and in Zea mays are described. Publicly available plasmid vectors that facilitate the stable chromosomal integration of inverted repeat transgenes that trigger RNAi have been used to generate more than 50 independent transgenic lines each in Arabidopsis and maize. Analysis of mRNA abundance of the targeted genes in independent lines transformed with distinct constructs indicates that the success of RNAi-induced silencing is gene dependent. mRNA levels were not detectably reduced for some genes, but were dramatically reduced for a number of genes targeted. A common pattern was that multiple independent lines transgenic for the same construct showed the same extent of silencing. This chapter describes the procedures used to generate and test transgenic lines mediating RNAi in Arabidopsis and maize.
authors.affiliation: Department of Plant Sciences, University of Arizona, Tucson, AZ 85721, USA.
authors.forename: Karen
authors.initials: K
authors.lastname: McGinnis
authors.forename: Vicki
authors.initials: V
authors.lastname: Chandler
authors.forename: Karen
authors.initials: K
authors.lastname: Cone
authors.forename: Heidi
authors.initials: H
authors.lastname: Kaeppler
authors.forename: Shawn
authors.initials: S
authors.lastname: Kaeppler
authors.forename: Arthur
authors.initials: A
authors.lastname: Kerschen
authors.forename: Craig
authors.initials: C
authors.lastname: Pikaard
authors.forename: Eric
authors.initials: E
authors.lastname: Richards
authors.forename: Lyudmila
authors.initials: L
authors.lastname: Sidorenko
authors.forename: Todd
authors.initials: T
authors.lastname: Smith
authors.forename: Nathan
authors.initials: N
authors.lastname: Springer
authors.forename: Tuya
authors.initials: T
authors.lastname: Wulan
mesh_terms: D017360:Arabidopsis / Q000235:genetics*; D018745:Genome, Plant; D023281:Genomics; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes; D003313:Zea mays / Q000235:genetics*