US 12,065,647 B2
Short/small hairpin RNA molecules
Elane Fishilevich, Indianapolis, IN (US); Kenneth Narva, Indianapolis, IN (US); Xiaozeng Yang, Indianapolis, IN (US); Meghan L. Frey, Indianapolis, IN (US); Murugesan Rangasamy, Indianapolis, IN (US); Wendy Lo, Indianapolis, IN (US); Premchand Gandra, Indianapolis, IN (US); and Peter Michael Waterhouse, Newton (AU)
Assigned to DOW AGROSCIENCES LLC, Indianapolis, IN (US); and QUEENSLAND UNIVERSITY OF TECHNOLOGY, Brisbane (AU)
Appl. No. 16/971,893
Filed by DOW AGROSCIENCES LLC, Indianapolis, IN (US); and QUEENSLAND UNIVERSITY OF TECHNOLOGY, Brisbane (AU)
PCT Filed Feb. 22, 2019, PCT No. PCT/AU2019/050146
§ 371(c)(1), (2) Date Aug. 21, 2020,
PCT Pub. No. WO2019/161449, PCT Pub. Date Aug. 29, 2019.
Claims priority of provisional application 62/773,355, filed on Nov. 30, 2018.
Claims priority of provisional application 62/633,720, filed on Feb. 22, 2018.
Prior Publication US 2021/0254058 A1, Aug. 19, 2021
Int. Cl. C12N 15/113 (2010.01); A01N 63/60 (2020.01); C12N 15/82 (2006.01)
CPC C12N 15/113 (2013.01) [A01N 63/60 (2020.01); C12N 15/8218 (2013.01); C12N 15/8286 (2013.01); C12N 2310/14 (2013.01); C12N 2310/531 (2013.01)] 12 Claims
 
1. A small hairpin RNA (shRNA) polynucleotide less than 70 nucleotides in length, the polynucleotide comprising the following structures:
a) a hairpin structure consisting of less than or equal to 25 nucleotides, obtained from an insect microRNA scaffold, that comprises double-stranded nucleotides and single-stranded nucleotides;
b) a DICER-1 recognition and cleavage site structure; and
c) a double-stranded stem structure consisting of 16-25 nucleotide pairs, wherein the two strands of the double-stranded stem structure are 100% complementary to one another;
wherein the two strands of the double-stranded stem structure (c) are connected at one end of the double-stranded stem structure via the hairpin structure (a); wherein the stem structure comprises a polynucleotide that shares 70% to 100% sequence identity with a target polynucleotide of a plant pest;
wherein the shRNA is unable to be cleaved by insect DROSHA.