Mutagenic inverted repeat assisted genome engineering (MIRAGE)

Nikhil U. Nair, Huimin Zhao

Research output: Contribution to journalArticlepeer-review


Here we describe a one-step method to create precise modifications in the genome of Saccharomyces cerevisiae as a tool for synthetic biology, metabolic engineering, systems biology and genetic studies. Through homologous recombination, a mutagenesis cassette containing an inverted repeat of selection marker(s) is integrated into the genome. Due to its inherent instability in genomic DNA, the inverted repeat catalyzes spontaneous self-excision, resulting in precise genome modification. Since this excision occurs at very high frequencies, selection for the integration event can be followed immediately by counterselection, without the need for growth in permissive conditions. This is the first time a truly one-step method has been described for genome modification in any organism.

Original languageEnglish (US)
Article numbere9
JournalNucleic acids research
Issue number1
StatePublished - 2009

ASJC Scopus subject areas

  • Genetics


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