Transient RNAi assay in 96-well plate format facilitates high-throughput gene function studies in planta.
Here we describe a rapid high-throughput method for performing RNA interference (RNAi) in moss, in which phenotyping is performed within 1 week after transformation. The moss Physcomitrella patens is a great plant model system for reverse genetic studies due to its amenability to homologous recombination as well as RNAi. Our lab has developed a rapid RNAi assay to screen for growth phenotypes in moss protonemal tissue. Here we describe how we have recently further facilitated this assay by modifying the PEG-mediated transformation protocol allowing for transformations to be carried out in a semiautomated fashion in a 96-well plate format.
Plant Biology Graduate Program, University of Massachusetts, Amherst, MA, USA.
Shu-Zon
SZ
Wu
Magdalena
M
Bezanilla
D019068:Bryopsida / Q000235:genetics*; D055785:Gene Knockdown Techniques / Q000379:methods*; D017343:Genes, Plant / Q000235:genetics*; D034622:RNA Interference; D014170:Transformation, Genetic