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Efficient gene targeting and removal of foreign DNA by homologous recombination in the picoeukaryote Ostreococcus
Author(s) -
Lozano JeanClaude,
Schatt Philippe,
Botebol Hugo,
Vergé Valérie,
Lesuisse Emmanuel,
Blain Stéphane,
Carré Isabelle A.,
Bouget FrançoisYves
Publication year - 2014
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/tpj.12530
Subject(s) - biology , homologous recombination , genetics , gene , gene targeting , transposable element , genome , functional genomics , computational biology , genomics
Summary With fewer than 8000 genes and a minimalist cellular organization, the green picoalga Ostreococcus tauri is one of the simplest photosynthetic eukaryotes. Ostreococcus tauri contains many plant‐specific genes but exhibits a very low gene redundancy. The haploid genome is extremely dense with few repeated sequences and rare transposons. Thanks to the implementation of genetic transformation and vectors for inducible overexpression/knockdown this picoeukaryotic alga has emerged in recent years as a model organism for functional genomics analyses and systems biology. Here we report the development of an efficient gene targeting technique which we use to knock out the nitrate reductase and ferritin genes and to knock in a luciferase reporter in frame to the ferritin native protein. Furthermore, we show that the frequency of insertion by homologous recombination is greatly enhanced when the transgene is designed to replace an existing genomic insertion. We propose that a natural mechanism based on homologous recombination may operate to remove inserted DNA sequences from the genome.