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Parental Rnai Suppression of Chromatin Remodeling Genes to Control Coleopteran Pests

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Parental Rnai Suppression of Chromatin Remodeling Genes to Control Coleopteran Pests University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Faculty Publications: Department of Entomology Entomology, Department of 2018 PARENTAL RNAI SUPPRESSION OF CHROMATIN REMODELING GENES TO CONTROL COLEOPTERAN PESTS Blair D. Siegfried Lincoln, NE, [email protected] Kenneth E. Narva Zionsville, IN Kanika Arora Indianapolis, IN Sarah E. Worden Indianapolis, IN Chitvan Khajuria Chesterfield, MO See next page for additional authors Follow this and additional works at: https://digitalcommons.unl.edu/entomologyfacpub Part of the Entomology Commons Siegfried, Blair D.; Narva, Kenneth E.; Arora, Kanika; Worden, Sarah E.; Khajuria, Chitvan; Fishilevich, Elane; Storer, Nicholas P.; Frey, Meghan; Hamm, Ronda L.; and Velez, Ana Maria, "PARENTAL RNAI SUPPRESSION OF CHROMATIN REMODELING GENES TO CONTROL COLEOPTERAN PESTS" (2018). Faculty Publications: Department of Entomology. 707. https://digitalcommons.unl.edu/entomologyfacpub/707 This Article is brought to you for free and open access by the Entomology, Department of at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Faculty Publications: Department of Entomology by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. Authors Blair D. Siegfried, Kenneth E. Narva, Kanika Arora, Sarah E. Worden, Chitvan Khajuria, Elane Fishilevich, Nicholas P. Storer, Meghan Frey, Ronda L. Hamm, and Ana Maria Velez This article is available at DigitalCommons@University of Nebraska - Lincoln: https://digitalcommons.unl.edu/ entomologyfacpub/707 |HAI LAMA ALTINA ATANAUS010053706B2NA MTOTO AU HATI (12 ) United States Patent (10 ) Patent No. : US 10 ,053 , 706 B2 Siegfried et al. (45 ) Date of Patent: Aug. 21 , 2018 ( 54 ) PARENTAL RNAI SUPPRESSION OF USPC .. 800 / 279 CHROMATIN REMODELING GENES TO See application file for complete search history . CONTROL COLEOPTERAN PESTS ( 56 ) References Cited (71 ) Applicants : Dow AgroSciences LLC , Zionsville , IN (US ) ; The Board of Regents of the U . S . PATENT DOCUMENTS University of Nebraska , Lincoln , NE 2009 /0188005 A1 * 7 /2009 Boukharov . .. C07K 14 / 4354 (US ) 800 / 279 2012/ 0174258 A1 7 / 2012 Narva et al. (72 ) Inventors: Blair D . Siegfried , Lincoln , NE (US ) ; 2013 / 0058890 A1 3 /2013 Raemaekers et al . Kenneth E . Narva , Zionsville , IN 2013 /0097730 A1 4 /2013 Narva (US ) ; Kanika Arora , Indianapolis , IN 2016 /0230186 AL 8 /2016 Baum (US ) ; Sarah E . Worden , Indianapolis , IN (US ) ; Chitvan Khajuria , FOREIGN PATENT DOCUMENTS Chesterfield , MO (US ) ; Elane WO 2007035650 3 / 2007 Fishilevich , Indianapolis , IN (US ) ; Nicholas P . Storer , Kensington , MD (US ) ; Meghan Frey , Greenwood , IN OTHER PUBLICATIONS (US ) ; Ronda L . Hamm , Carmel , IN Thomas et al . 2001 , The Plant Journal 25 ( 4 ) :417 -425 . * (US ) ; Ana Velez , Lincoln , NE (US ) Yibrah et al . 1993 , Hereditas 118 :273 - 2890 . * Thomas et al ., 2001 ; Plant J . 25 :417 -425 . ( 73 ) Assignees: Dow AgroSciences LLC , Indianapolis , Gkhajuria , Chitvan et al ., ' Parental RNA interference of genes involved in embryonic development of the western corn rootworm , IN (US ) ; The Board of Regents of the Diabrotica virgifera virgifera LeConte ', Insect Biochemistry and University of Nebraska , Lincoln , NE Molecular Biology , Epub . May 22 , 2015 , vol. 63 , pp . 54 -62 . (US ) Ho , Lena et al. , “ Chromatin remodelling during development, ” , Nature, Jan . 28 , 2010 , pp . 474 -484 , vol. 463 , No . 7280 (NIH Public ( * ) Notice : Subject to any disclaimer , the term of this Access Author Manuscript Version internal pp . 1 - 24 ) . patent is extended or adjusted under 35 Mito , Taro et al ., “ Kruppel acts as a gap gene regulating expression U . S . C . 154 ( b ) by 92 days. of hunchback and even - skipped in the intermediate germ cricket Glyllus bimaculatus, ” Developmental Biology, Epub . Apr. 17 , (21 ) Appl. No. : 14 /971 ,515 2006 , vol. 294 , No . 2 , pp . 471 -481 . NCBI, GenBank accession No. XM _ 003742362 . 1 ( Jun . 14 , 2012 ) . NCBI, GenBank accession No . XM _ 007547345 . 1 ( Apr. 22 , 2014 ) . (22 ) Filed : Dec . 16 , 2015 Search Report and Written Opinion for PCT/ US2015 / 066082 , dated Apr . 15, 2016 (65 ) Prior Publication Data Search Report and Written Opinion for PCT/ US2015 /066134 , dated US 2016 / 0222408 A1 Aug. 4 , 2016 Apr. 22 , 2016 . Related U . S . Application Data * cited by examiner ( 60 ) Provisional application No . 62/ 092 ,747 , filed on Dec . Primary Examiner - Li Zheng 16 , 2014 . ( 74 ) Attorney , Agent, or Firm — Sean M . Russell; Magleby Cataxinos & Greenwood (51 ) Int. Cl. C12N 15 /82 ( 2006 . 01 ) (57 ) ABSTRACT C12N 15 / 113 ( 2010 . 01 ) This disclosure concerns nucleic acid molecules and meth AOIN 57 / 16 ( 2006 . 01 ) ods of use thereof for control of hemipteran pests through AOIN 63 /02 ( 2006 .01 ) RNA interference -mediated inhibition of target coding and (52 ) U . S . CI. transcribed non - coding sequences in hemipteran pests . The CPC . .. .. .. C12N 15 /8286 (2013 . 01 ) ; AOIN 57 / 16 disclosure also concerns methods for making transgenic (2013 .01 ) ; AOIN 63 /02 ( 2013 . 01 ) ; C12N plants that express nucleic acid molecules useful for the 15 / 113 (2013 .01 ) ; C12N 15 /8218 ( 2013. 01 ) ; control of hemipteran pests , and the plant cells and plants C12N 2310 / 14 (2013 . 01 ) obtained thereby . (58 ) Field of Classification Search ??? C12N 15 /8286 31 Claims, 11 Drawing Sheets U . S . Patent Aug . 21, 2018 Sheet 1 of 11 US 10 ,053 , 706 B2 | * 3 2011:11 Wii 2000W OOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOO! 0000000000000000000?????????????06dddddddddddddddddddddddddd; dsRNA wiiiiiiiiiiiiiiiiiiw OSRNA ooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooo .CCCCCCCCCCC000000000OOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOO ooooooooooooooooooooooooooooooooooooooooooooooooooooooooooo11 -0000000000000000000000000000000000000000000000000000000M WWW winninn ' :W wwwwwwwwwwwwwwwiiiiiiiiiiiiin W WW ?????????????????????????????????????????????????????????? WWW W ????????????????????????????????????????????????? oooooooooooooooooooooooooooooooooW 0000000000000000000000000000000000000000000000000000000000000 ???????????????????????????????????????????????????????????? Woooo00000000000000000000000000000000000000000000000000OOOO W90.2 FIG.1A 44444444444 FIG.1B *. 17RNA polymerase T7RNA polymerase 37°C14h Template1 00000000000000000000000000000000000000000000000000000000000 Template2 U . S . Patent Aug . 21, 2018 Sheet 2 of 11 US 10 ,053 ,706 B2 { ???????? Chdi WCR 1001 Chd1 Eheros 100 - Chd1 D .melanogaster - Mi- 2 D .melanogaster 100 www. Mi- 2 E heros 521 M2WCR SNF2 S .cerevisiae BRAHMA H sapiens 100 - Brahma WCR 100 Brahma E heros 100 - Brahma D melanogaster IsW - 3 WCR 70 5W -1 WCR 100 L . (sWi - 2 WOR 857mm sw D .melanogaster 99 r sw - 1 E heros 55 [liswi - 1 H.: halys 100 sw - 2 E heros 96 1sWi- 2 H hays 02 FIG . 2 U . S . Patent Aug . 21, 2018 Sheet 3 of 11 US 10 , 053 , 706 B2 ** w 1.4,5*.* SLIDESANT-HANDcase bromodomain DUF4208 * * * * * * * * helicase * * tekcase * = S FIG.34 FIG.3B helicase FIG.30 . 1. moms 2 2.1 - . SIA BRKQLQ Isw-1 IsWi-2 15W-3 - Iswi-2 SWI2/NSF2family:brahma . ISWIfamily:IswiDBINO Chromodoma co WCRbrahma, aneseconservers.wowwww CHDsubfamily1:Chdl BSBbrahma Diebrahma BSBISW WCRISWI Dmelw Chi Wwwwwww BSBCdi DmChat U . S . Patent Aug . 21, 2018 Sheet 4 of 11 US 10 ,053 , 706 B2 CHIEN DUTOS 0092086 * DUF1087 helicase helicase * FIG.3D 3E.FIG BE 3892 CHDsubfamilyII:Mi-2 chromodomainPHDCHDNT CHDsubfamilyIII:kismet chromodomain -1114 WCRMi-2 BSBMi-2 DmeMi-2 WCRkis BSBkis Dmelis* * U . S . Patent Aug . 21, 2018 Sheet 5 of 11 US 10 , 053, 706 B2 week1 week2 -2 4 iswi-2 ** chd1SWK. * FIG.4B brm Oviposition/dayfemale emaniswi-1 2iswi-men m *YFP neworm neichd1 m - gram r p 20 magelse E 1015 minem aprengerpartypropongonggongnanganggopaggagawaangforgangsprogramangng dayspostinjection FIG.4A 5 20anosKwania home funnanborrannmandenmannnnndwannananandannawanaendanamanwand 0 #Survivingfemales die u o U . S . Patent Aug . 21, 2018 Sheet 6 of 11 US 10 ,053 , 706 B2 notinjected dsRNA-injected W quo chdi mi-2iswi1 dsRNA ?.??pyars } wweek1 mweek2 FIG.5 1-2 † Relatvexpress in expression Relative swi-2 † ísWi- TILLE † chd? FIG.40 †* brm Eggshatched'day,female me . o . o - U . S . Patent Aug . 21, 2018 Sheet 7 of 11 US 10 ,053 ,706 B2 days1-3 5210.days wdays7.9 1813-daysen wdaya. FIG.68 OSRNA Ih YED eggshatchedperfemale wwwmmmmmmmmmmmmmmmmnnnnnnnnnnnnn days1-3days6 79days days10.12days13-15 w 2mon * try i . www frumoase dsRNA YEP www FIG.6A perfemalelaideges U . S . Patent Aug . 21, 2018 Sheet 8 of 11 US 10 ,053 , 706 B2 4-dayold FIG7B 14dayspostinjection FIG7D 0-dayold FIG7A 9dayspostinjection FIG7C VÁ YEP DARNA injectedun- VIS U . S . Patent Aug . 21, 2018 Sheet 9 of 11 US 10 ,053 , 706 B2 2mm FIGZF 14dayspostinjection FIG7H 14dayspostinjection FIG7E 9dayspostinjection 9dayspostinjection FIG7G dsRNA SEMmin? ASRNA bralma U . S . Patent Aug . 21, 2018 Sheet 10 of 11 US 10 , 053 , 706 B2 1 0.9 overRallelewithoutpRNAI stensomYallele OXXsecondo poco con000 RelativemagnitudeofthePRNAieffectonfemaleB$adultsemergingfromrefugepatch OOOO.OOOOO .* *. * * . * * FIG.8 ***** 0.12345678 medsort or v O se* 4 STE &#6 -- o&# KO O& .. + foto +*. OO eria #. .5freq??cyfaces3 s 0 Generationsuntilallele 300 100 50 O U . S . Patent Aug . 21, 2018 Sheet 11 of 11 US 10 ,053 ,706 B2 1 0.9 -YYYYYYYYYYYYYYYYYYYYYYYYYYY oooooooooooooooooooooooooo **** ** azerinisRalleleandYallelewithoutpRNAI o e **** *Rallele *Yallele *** de oro enoto * 0.8764530.2
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