Description:
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The GE-PPO Mushroom is a transgene-free, anti-browning white button mushroom (Agaricus bisporus). In this mushroom, small deletions (1 to 14 bp) were introduced by CRISPR/Cas9 in one of the six polyphenol oxidase gene (ppo).
PPO is responsible for the enzymatic browning (melanin formation). ppo knock-out leads to a reduction of the enzyme's activity and thereby to the mushroom’s anti-browning property which improves the appearance and shelf life. This is expected to facilitate automated mechanical harvesting.
A pUC19-based construct carrying the gRNA and the Cas9 was introduced into mushroom protoplasts by PEG-mediated transformation and was transiently expressed. No selection marker was used.
GE-PPO Mushroom strains containing no foreign DNA integration in its genome, as verified by PCR and Southern blot analyses, were selected for further studies and potential commercialization.
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Links regarding to this GMO:
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USDA-APHIS http://www.aphis.usda.gov/biotechnology/downloads/reg_loi/15-321-01_air_inquiry.pdf
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Transformation / Modification technique:
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CRISPR/Cas
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