Deleteagene™ (Delete-a-gene) is a deletion-based gene knockout system for plants. To
obtain deletion mutants for a specific gene, random deletion libraries created by fast
neutron mutagenesis are screened by polymerase chain reaction (PCR) using primers
flanking the target gene. By adjusting the PCR extension time to preferentially amplify the
deletion alleles, deletion mutants can be identified in pools of DNA samples with each
sample representing more than a thousand mutant lines. In Arabidopsis, knockout plants
for greater than 80% of targeted genes have been obtained from a population of 51 840
lines. A large number of deletion mutants have been identified and multiple deletion alleles
are often recovered for targeted loci. In Arabidopsis, the method is very useful for
targeting small genes and can be used to find deletion mutants mutating two or three
tandem homologous genes. In addition, the method is demonstrated to be effective in rice
as a deletion mutant for a rice gene was obtained with a similar approach. Because fast
neutron mutagenesis is applicable to all plant genetic systems, Deleteagene™ has the potential to enable reverse genetics for a wide range of plant species.
