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Comparative and Functional Genomics
Volume 3, Issue 2, Pages 158-160
http://dx.doi.org/10.1002/cfg.148
Conference Review

Deleteagene: A Fast Neutron Deletion Mutagenesis-Based Gene Knockout System for Plants

1Biotechnology Laboratory, Room 237, 6174 University Blvd., University of British Columbia, V6T 1Z3, BC, Canada
2Maxygen, Inc., 515 Galveston Drive, Redwood City 94063, CA, USA

Received 29 January 2002; Accepted 30 January 2002

Copyright © 2002 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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.