Replication Independent Formation of Extrachromosomal Circular DNA in Mammalian Cell-Free System 英文参考文献.docVIP

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Replication Independent Formation of Extrachromosomal Circular DNA in Mammalian Cell-Free System 英文参考文献.doc

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Replication Independent Formation of Extrachromosomal Circular DNA in Mammalian Cell-Free System 英文参考文献

ReplicationIndependentFormationof ExtrachromosomalCircularDNAinMammalianCell-Free System ZoyaCohen,SaraLavi* DepartmentofCellResearchandImmunology,TelAvivUniversity,TelAviv,Israel Abstract ExtrachromosomalcircularDNA(eccDNA)isapoolofcirculardoublestrandedDNAmoleculesfoundinalleukaryoticcells and composed of repeated chromosomal sequences. It was proposed to be involved in genomic instability, aging and alternative telomere lengthening. Our study presents novel mammalian cell-free system for eccDNA generation. Using purifiedproteinextractweshowthateccDNAformationdoesnotinvolvede-novoDNAsynthesissuggestingthateccDNAis generatedthroughexcisionofchromosomalsequences.Thisprocessiscarriedoutbysequence-independentenzymesas human protein extract can produce mouse- specific eccDNA from high molecular weight mouse DNA, and vice versa. EccDNAproductiondoesnotdependonATP,requiresresidualamountsofMg2+andisenhancedbydoublestrandDNA breaks. Citation:CohenZ,LaviS(2009)ReplicationIndependentFormationofExtrachromosomalCircularDNAinMammalianCell-FreeSystem.PLoSONE4(7):e6126. doi:10.1371/journal.pone.0006126 Editor:BethA.Sullivan,DukeUniversity,UnitedStatesofAmerica ReceivedMarch19,2009;AcceptedMay29,2009;PublishedJuly1,2009 Copyright: ? 2009 Cohen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:ThisresearchwasfundedbytheIsraelCancerAssociationthroughtheestateofthelateEthelCohen.Thefundershadnoroleinstudydesign,data collectionandanalysis,decisiontopublish,orpreparationofthemanuscript. CompetingInterests:Theauthorshavedeclaredthatnocompetinginterestsexist. *E-mail:lavisara@post.tau.ac.il Introduction ThecurrentstudyisaimedtoexplorethefirststepsofeccDNA synthesisinmammaliancells.Tothisendwehaveconstructeda mammalian cell free system. Using this system we show that eccDNA formation from high molecular