Journal
NATURE BIOTECHNOLOGY
Volume 36, Issue 9, Pages 894-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nbt.4202
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Funding
- National Key Research and Development Program of China [2016YFD0101804]
- National Natural Science Foundation of China [31788103, 31420103912, 31501376, 31570369]
- Chinese Academy of Sciences [QYZDY-SSW-SMC030, GJHZ1602]
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Translational regulation by upstream open reading frames (uORFs) is becoming established as a general mechanism for controlling the amount of protein that is synthesized from downstream primary ORFs (pORFs)(1-5). We found that genome editing of endogenous uORFs in plants enabled the modulation of translation of mRNAs from four pORFs that are involved in either development or antioxidant biosynthesis. A single-guide RNA that targeted the region harboring a uORF initiation codon can produce multiple mutations. Following uORF editing, we observed varying amounts of mRNA translation in four pORFs. Notably, editing the uORF of LsGGP2, which encodes a key enzyme in vitamin C biosynthesis in lettuce, not only increased oxidation stress tolerance, but also increased ascorbate content by similar to 150%. These data indicate that editing plant uORFs provides a generalizable, efficient method for manipulating translation of mRNA that could be applied to dissect biological mechanisms and improve crops.
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