学科分类
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4 个结果
  • 简介:Precisebaseeditingishighlydesiredinplantfunctionalgenomicresearchandcropmolecularbreeding.Inthisstudy,weconstructedarice-codonoptimizedadeninebaseeditor(ABE)-nCas9toolthatinducedtargetedA·TtoG·Cpointmutationofakeysinglenucleotidepolymorphismsiteinanimportantagriculturalgene.Combinedwiththemodifiedsingle-guideRNAvariant,ourplantABEtoolcanefficientlyachieveadeninebaseeditinginthericegenome.

  • 标签: PRECISE optimized ADENINE AGRICULTURAL GENE
  • 简介:EliminationoftheCRISPR/Cas9constructsineditedplantsisaprerequisiteforassessinggeneticstability,conductingphenotypiccharacterization,andapplyingforcommercializationoftheplants.However,removaloftheCRISPR/Cas9transgenesbygeneticsegregationandbybackcrossislaboriousandtimeconsuming.WepreviouslyreportedthedevelopmentofthetransgenekillerCRISPR(TKC)technologythatusesapairofsuicidegenestotriggerself-eliminationofthetransgeneswithoutcompromisinggeneeditingefficiency.TheTKCtechnologyenablesisolationoftransgene-freeCRISPR-editedplantswithinasinglegeneration,greatlyacceleratingcropimprovements.Here,wepresentedtwonewTKCvectorsthatshowgreatefficiencyinbotheditingthetargetgeneandinundergoingself-eliminationofthetransgenes.ThenewvectorsreplacedtheCaMV35SpromoterusedinourpreviousTKCvectorwithtworicepromoterstodriveoneofthesuicidegenes,providingadvantagesoverourpreviousTKCvectorundercertainconditions.Thevectorsreportedhereofferedmoreoptionsandflexibilitytoconductgeneeditingexperimentsinrice.

  • 标签: genome EDITING SUICIDE gene TRANSGENE KILLER
  • 简介:RiceisanimportantfoodcropinChina,andthedevelopmentofhybridriceisacrucialwaytoincreasegrainyield.Thecreationofdual-purposenuclear-sterilelinesfortwo-linehybridbreedinghasbecomevitalforcommercialricebreeding.WeconstructedthepC1300-2x35S::Cas9-sgRNAPTGMS2-1expressionvectorforeditingthemalefertilitygenePTGMS2-1intwowidelycompatiblericevarieties,93-11andHuazhan,byusingtheCRISPR/Cas9system.Weobtainedthemarker-freephotoperiod-/thermo-sensitivegenicmale-sterile(P/TGMS)linesinT1generation.Accordingtotheexperimentsinphytotronwithfourtemperatureandphotoperiodtreatments,wefoundthetemperatureisthemainfactorforrestoringthepollenfertilityofptgms2-1mutantsin93-11andHuazhan,andthephotoperiodalsohassomeeffectsonpollenfertilityintwodifferentricebackgrounds.Theapplicationofcultivatingnewmale-sterilelinesbygenomeeditingsystemwillsignificantlyacceleratethericebreedingprocess.

  • 标签: RICE CREATION MARKER-FREE
  • 简介:Homeoboxtranscriptionfactorsparticipateinthegrowthanddevelopmentofplantsbyregulatingcelldifferentiation,morphogenesisandenvironmentalsignalresponse.Torevealthefunctionsofthesetranscriptionfactorsinrice,weconstructedtheRNAivectorsofOsHox9,amemberofhomeoboxfamily,andanalyzedthefunctionofOsHox9usingreversegenetics.TheplantheightandtilleringnumberofRNAitransgenicplantsdecreasedcomparedwiththoseofwild-typeplants.Reversetranscription-polymerasechainreactionanalysisshowedthatOsHox9expressionreducedinthetransgenicplantswithphenotypicvariance,whereasthatinthetransgenicplantswithoutphenotypicvariancewassimilartothatinthewild-typeplants.ThisresultsuggeststhatthephenotypesofthetransgenicplantswerecausedbyRNAieffects.Thetissue-specificityofOsHox9expressionindicatedthatitwasexpressedindifferentorgans,withhighexpressioninstemapicalmeristemandyoungpanicles.SubcellularlocationofOsHox9demonstratedthatitwaslocalizedonthecellmembrane.

  • 标签: 转录因子 同源盒 遗传分析 家族 水稻 转基因植物