title: A novel method of demonstrating the molecular and functional equivalence between in vitro and plant-produced double-stranded RNA.
abstract: A biotechnology-derived corn variety, MON 87411, containing a suppression cassette that expresses an inverted repeat sequence that matches the sequence of western corn rootworm (WCR; Diabrotica virgifera virgifera) has been developed. The expression of the cassette results in the formation of a double-stranded RNA (dsRNA) transcript containing a 240 bp fragment of the WCR Snf7 gene (DvSnf7) that confers resistance to corn rootworm by suppressing levels of DvSnf7 mRNA in WCR after root feeding. Internationally accepted guidelines for the assessment of genetically modified crop products have been developed to ensure that these plants are as safe for food, feed, and environmental release as their non-modified counterparts (Codex, 2009). As part of these assessments MON 87411 must undergo an extensive environmental assessment that requires large quantities of DvSnf7 dsRNA that was produced by in vitro transcription (IVT). To determine if the IVT dsRNA is a suitable surrogate for the MON 87411-produced DvSnf7 dsRNA in regulatory studies, the nucleotide sequence, secondary structure, and functional activity of each were characterized and demonstrated to be comparable. This comprehensive characterization indicates that the IVT DvSnf7 dsRNA is equivalent to the MON 87411-produced DvSnf7 dsRNA and it is a suitable surrogate for regulatory studies.
authors.affiliation: Monsanto Company, 800 North Lindbergh Blvd, St. Louis, Missouri 63167, United States. Electronic address: william.urquhart@monsanto.com.
authors.forename: William
authors.initials: W
authors.lastname: Urquhart
authors.affiliation: Monsanto Company, 800 North Lindbergh Blvd, St. Louis, Missouri 63167, United States.
authors.forename: Geoffrey M
authors.initials: GM
authors.lastname: Mueller
authors.affiliation: Monsanto Company, 800 North Lindbergh Blvd, St. Louis, Missouri 63167, United States.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Carleton
authors.affiliation: Monsanto Company, 800 North Lindbergh Blvd, St. Louis, Missouri 63167, United States.
authors.forename: Zihong
authors.initials: Z
authors.lastname: Song
authors.affiliation: Monsanto Company, 800 North Lindbergh Blvd, St. Louis, Missouri 63167, United States.
authors.forename: Tim
authors.initials: T
authors.lastname: Perez
authors.affiliation: Monsanto Company, 800 North Lindbergh Blvd, St. Louis, Missouri 63167, United States.
authors.forename: Joshua P
authors.initials: JP
authors.lastname: Uffman
authors.affiliation: Monsanto Company, 800 North Lindbergh Blvd, St. Louis, Missouri 63167, United States.
authors.forename: Peter D
authors.initials: PD
authors.lastname: Jensen
authors.affiliation: Monsanto Company, 800 North Lindbergh Blvd, St. Louis, Missouri 63167, United States.
authors.forename: Steven L
authors.initials: SL
authors.lastname: Levine
authors.affiliation: Monsanto Company, 800 North Lindbergh Blvd, St. Louis, Missouri 63167, United States.
authors.forename: Jason
authors.initials: J
authors.lastname: Ward
mesh_terms: D000818:Animals; D001517:Coleoptera; D004305:Dose-Response Relationship, Drug; D007300:Insect Control / Q000379:methods; D010572:Pest Control, Biological / Q000379:methods; D018517:Plant Roots / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000378:metabolism; D012330:RNA, Double-Stranded / Q000096:biosynthesis / Q000235:genetics*; D003313:Zea mays / Q000235:genetics* / Q000378:metabolism
chemical_list: D012330:RNA, Double-Stranded
keywords: Double stranded RNA; DvSnf7; Functional equivalence; MON 87411; Molecular equivalence; Prediction interval; Safety assessment; dsRNA