Nuclear gene targeting in Chlamydomonas reinhardtii

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By means of a model gene we have shown for Chlamydomonas reinhardtii that single stranded DNA is able to target or modifiy nuclear genes more efficiently than double stranded DNA. In addition, non-homologous DNA-Integration is nearly completely suppressed. These findings open the possibility of silencing or modification of any selected gene in green algae. During the next three years we intend to optimize this "Nuclear Gene Targeting System" and make it applicable especially for weakly expressed genes. To increase the recombination efficiency we will over-express some genes that are involved in recombination processes from Streptomyces rimosus, Ps. aeruginosa or Chlamydomonas itself or add purified recombination proteins to ssDNA during transformation.