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This mutant showed PT weight and exhibited no growth problem or auxotrophy. The thiI gene had been more examined for the use as a range marker in genome co-editing. Ribonucleoprotein complex comprising recombinant Cas9 nuclease and sgRNA focusing on thiI or any other target gene (wA or sreA) had been ready and simultaneously introduced into A. oryzae RIB40. thiI and target gene dual loss-of-function mutants had been effortlessly selected on PT-co