Enhanced tnpb polypeptide and use in genomic editing

Abstract

In one aspect, the invention relates to a TnpB variant polypeptide comprising a Deinococcus radiodurans (ISDra2) TnpB polypeptide sequence and, separated by a peptide linker, a nuclear localization module comprising at least two nuclear localization sequences (NLS) separated by a first peptide linker. A second aspect of the invention relates to a DNA editing polypeptide comprising the TnpB variant polypeptide according to the invention, wherein the TnpB variant polypeptide comprises a nuclease deactivating mutation, and a nucleoside deaminase. The invention further relates to a nucleic acid sequence encoding a polypeptide according to the invention, and to a virus comprising the nucleic sequence.

Description

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rrnr nu ’,’ m nr n . rvr nn m rn rzn un, rzn n m m rm v nrmn,r mxrrrrr. n r n nrznnunnxnn nx rn m mmmnn,r rvrn-mmvnr rrr nn. wn ,r mrznm n mm r n nn-mn n . mnmn r , nrrmnnnrnrnn n m,nn, wn r n r n mr.nrnn mr mrm, nu xr n vrn nx rn n rrn vrnmrn , w rn n mrn r n nn vrnm r w rnnnr,r -n rn mr.rn vrnr r-nrnnv rrrnrnnnnrnnnrrm n nr nrnr rmr nn r rn n, rnn r ,, nnr rnr rmnr n rn m r nnn ’ .nrnmm vrn n m vr vr, w rn n,r vr.nm rnrm m nnnx rn n r n nn n n rnr n nr r m. mn.mr m n v “mnr nrnrn rn’ nnn nr n r m nnn rn. r mn. m mr ’ -m nnn ’ r rn r mnvnnnv rnn n,n vrn mrn,rnn, - nn u r urnnr nrm : nr , -n nnr n u r urn r n n n rn nznr mzmrnr n r zn n n r nn n, r n r rm nnr. n rr.rm n r zn m nnrunrmznun nmn.,nv n rrzn m -rmn n n nrvn nr v nrnnxmrnm. rmn, mn nrr zn m mr: m.nrrn nrnrnrr mrmnnmmnnr,n nnnrrn.zn m mr r .nr rnrnnm,mnr :. n nr,nr nnnrrnmn n,r r wrnnvnr m m m . xm v n w n wnnr r vrnnrnm rmn,v n rnrn. mn n.nr r mmmnr,mrn nnrnn n: . n n r zn vn vvnrr rmmnn,n n r z:n mnvn :,r n rn:, , wnn-rmnn v :. rmmmrn, n n nr :m n znnzn mn nr nx nr m rwnr zm n . rrr r rm wn:v vm v m v n m rrnrnmmn nvnn r. n vrn n mrn, :, rr vr, rnvnn vnn,rvnn,rnr vn:,n, mr vrnv n vn: n nx v :. v v n w v mn n n n nxm: r w rn rmn rn n nmr n rnn ,,nm .rmnmnrmn mrm .m wn :.rnw mn,.nn rrrnrnnnm,,.nnn.r r rrn rm nn-rm m mnn rrn rrrnrr .r. -mrn. mun, nn.n rrm rv rmx,rm rrm r, r rn mnrwrn rvr rmnn.mr nrr wnrmr,nnn -    rx rn mn rr’ x n n n .nvrnnr nrn. n rv mn, , mn.rrr n n mn . r-n.mn n rnz . “r-m rrnrnnn,–rrn”n r nn mrnmn, rmvn, n v:rnr :r, rn vn,vnnnrrnz, vnnxmn,n,xnn, nnr,nn, rnr rnn.nr rmmn,n m, mn n.n.nr, n .nr rmm rrrrmmmmnn,, m m nnn vr.nr rmmn, m n . nrr rmnvmnn, m n . rn nnn r n mrn n vrn n vr nmmn rn, wrn mn.n mr n vn m n,n n -mn nvzmn-nmmnnnmnnnrr:mmnr,mrrn nmm : -nr: mnn r mrmn m nnzm, - r: - mm r rrm nn mnr:nnzm, - r: - - vnrnnmrn: r: r . nr: - - n - nr nnmnmnrr:n nnrr:n r.:mnmr: nr w r n mn nnvn: mmnnrnwnnr r xn nrv r r :-,r r xn r. n .,nrnmmn, n ,nvnw m.n, r m nnr , r:v-.nmm:m.nn,r nrmmnm,n nnnnvnm m n .nrnmmn, nn mn n n. nzm n r : .nrnm xm. v mn, n mn -rmn rv nnrnr .nxm r nrn :. vrnnmm.nn,x nm r mnnrn -rm:n r.v n vnnnrrmrmmnn,r n mn n n vrn n mrr rmm. nvn mrr rmnm,nn,rmnrm nrnr mnmnnn. n. nnnrmnrmmn,nn rrm.nxmnrnnr :n r xm n rn n . nmn vnnrm nr nnvnn rrm n: nn :. rn nvnn.nr nn n vrnnn n nv rn nvnnn r n nn.r rnmmzmnn, nr mn vnnn.mzrrnrnnmnxr nn rnm. n n n nmrv mzn n nx rnrnnmn , n . nvv rn n n m rrr n mn r nnn nn r rnrn n, n rv - nnxr:n.rnrnnmvnrmnzrnrrnmrn n : mmn . nmzn nnrnn n n mn , w nmrvrn n n. n w n - wnv : nn m rnz nn vr rn n . n n rrn nn nmn , - nn:r nnm mzn mr n. m nn mmnnnv, wv n nn n . nnnrnrnn m - m m rm:nnrmznn mrv m vn n -vn rrrrrn r nnmn . rn nr mn: mznr n rmvr n nn, m r nnn r rmn, , -nnmn mrnrn r mnr . nn r mn nn: mzn n n vr n . nn mvmnn m n mr nrn v rnrr mnnmznn nvnrr rnxr, rmznn nnr .mnrvrnn rr n nn nn n nn, r nnmrvrn rnr. nrn n r m rnn nnr r rn nnvvnn nn. n r, nxr n vrmrn n n r v w,nnrn n vrn r mmmn,r rn r n mr r nnr rmmn, xrnr mr. nnnxr nrnnn vrrmrn rmrrnrrnxr nn rmr,.nnrmnrmmrnm,rxr n mmmn r rn : . xr rn mrnn w r rm xm rmr,n w n rmr,r w n,n rzm xm: ,n’ : rmnnn rnn ’ ’rnn. nnxrr nn6vnnnn r m vrn. n rmrn n nnn xr,n v vr r n rrrn n ,r vr r rv rr.nr rmmn mr n vr r rn - vr. mr mn rn nn r n m n. m vr r rnnrnn, n,n n n r. nn,nxr n, vrr mnx n wmnnmrn , w wnrnnnrnmnm nn r . .vnnnrrnm wnmmn: n vrn mnrn,rnn, -nn u r urn r nrm : nr , -nnr n u r urn r n v n n rn nznr mzmnrnn,r nrr zn n n r n r rm nn n n nnrr rm n r zn m. n r. zn v mrn -rrmnnmnmn, wrn nn.r. znn v mrn mr nrn mmn:r, w.rn n r. wrnn v nrnr zn mrnmnn r nmmn,. n vrn rn mrmnr, wrn. r. n v:rn. wrn r nrrnmnnnn,rrnr wmrnmn, n wr vrrnn nn n. n rnrn mnn. r nmmn, nn nnrrnmrr mnn n,r r: wr.n. wrnn v nrnr zn mrn nn:.r nmmn,. wrnn vrn rn nn r nmmn, rm n n nr : m rnnnz mn nr nx .n vrn nzrn zm n . :, mrn,rnn, rr vrrnvnn vnn,rvnn,r r vn n, mr vrn n :, . n vn v :. rnrz vrn m n, rvrn nn: rr nm:mn, r v, n n,n nn, n , rnn, rnr rnn. , n v. nr vrn rn mmn, wrn m n. m r , n . .n vrn rn mmn, wrn m n. - nn v mrn wrnrnn vrn rnmrnn n r nvmnmn, n m n,. - n vn mn mn. n rn mmn, wrn n wr m nn , rv :,r r. n nn vnr mnnm . mn nn mn r, wrn n.mn n n mrnn.mmn, wrn nn. n nr : . n rn nn mmn , wrn. nmn r-nrmnn rnvmnmn vrn, wr. n mn -rmn rv n v n. n rn nn mmnrn . n mn n n vrn , wrn nr nnrm.. nrn mnrn mmn, wrnnrm n nn,r r wrnnrm . r nmn n mn n rrnnn mn.n:mmn ,nn. n nnn n vrn n :. ,r n rn rn nn mmn. n n rn m nn mmn . n nmzrr mn, wrn n.rnxr n vrmrnn nrnnmnn.rn mmn nr,nnrrnr n mrmr. r n mmmn,r r v .xrxrnn vrnnrnnxmr mnn,rrrnrmrnrn mmmn mrr n . r rn rmr. mxr n vr rn mmn, wrn xr . vrrmnr wn nr rmn : . ,r xr n vr rn rn nn mmn r . vr r rn mmnn,n mmn . . m rn nn r n wrn vr n vr. mnrnnn , n mnn, m mrn vr r n r nmm ,nxr n vrr rvrrnvrn r n, nn n r. nr rnv m mrr rn “mmn”,nvn nr rmrmmn mnvn mnnnnrr vnrn.rnrwn.wnxxmmrn m r,rm w rr n m . n r nvnn .rn mvur rrnn nn mmmn wnn nmx. wr . rnn nnn r rn r nnn nn nnn n.n nn n rnn mn mnn nr’rmmnrn ωn rvnrm r n -rn n - n m . mrnn rxn,m rnnm, r. m,mn , r. nm nn mnmrn nnnn , r wr rn ω nn.n,nrnn vr . nmrn n n, rrn n -n . n = mn n =nnn r .r rrn mn ,nrnn-nrnrn-nn.-, nr r z,nn-nrn. ,nmr =mnn anmnn rnn rn n .r rrn. mnn n rrnn mnnn nr .vn-vmrznnnnrnmn ..n..mxmmnr =n., v = mmx, mmnn = =..,, = = mmmxmmnn = = a..,, = = rmmn =m.n, =., mn =., = rrn mn n n..n, nmrnv r . rrnn n - nn n nn r r.n nnm nn vnn-v.nnr wnn - - r - n.nn rr n-n r wnnvr .nn r rr-mr r. v r rrrnn rv. m rrnn r-m ωrnrrr ω n. ,rn n n m , mm r, rn vrn r . rn n, . r- n n rrnn zmxn, v rn rmr rn: :,nr r rrmn rw rn rmr rn: :,nn rn: : : vr. n rn rn r n nr-r w rn rrnnnnnnrn nrn w r v ω. xr xrnnrrn r .- n-nnm n rrn mn n =n vrn mm n ω v. . rmzn-m nnn , rnn =nv r n nr r rn. n =nnn r r w.n- nn ω n - nn ω v rv n ω., nmr mnnv =.,r = -n-mnmn =n. =,. =, =--nmn =., = -n =., = -nmn =., = -nmnn =m.n, = =., =-nm-nn =mn = -nmn =., =. m vrvw ., n -r n. , -rn xn - wr,w-r rrnnrnn rn n-rn’ n -. n : r: ’rn -r: -r: _ : -r: : nr.: w-:v :wnn mm: n-r : - r nr wnn :. rr- rnn r. rrzn -nnrn nn v rn-v . r .,-r - nn. v r rrwnv ω nn v r n. : nrmn vn mm vnnv nn n wnznwr vnn ’- . nn- rrn vr n =nnn r .n rr.r n ’-r .n r-nnrm m. rv n n- nmxn rn nnr vrn r.- rm- vrn r n’- ,n’- nv nm.xn nnn r . n- n = r n nn- vnnv mn n ’- ,’- ,’- ,’- . rrn n =-n.nnnr rn n.n rn -r rn r: : nn’’ r: : w v wr: : nrr n mm n-r : - :. : - : r : . rrmm n vvn - r nr wnn r. rn’nnmvn wnmx. m rrnn n, n . n nrn ω.v, n mn,mn n =nnn r mnm ω rrvrmmnn w vr rzm .r-nmvrn ω wn w n =nn nnv r n xrm n r . r nn rnn ω v. r- r nm-rn ω n r - m rrnnrrwnn ω rn vv v n. nrn vv . ,-nr-n n -mmnr rmr,vr- rmr, - vr r ,- r , n r zn n r, nnrmrr ω xr n vr rnrn r r vrm rnz.m m, rnrrnn n n r n nwrnr m. , mrnrvnrr.-n m n nm rmn r.nn vn r rnn rn. mr w. v m , m , mrn,,rnrm ,,rx , m ,nr. rrn rmnnm r rrm, mn.. n =nm. nn nwrn m rn r. v -n- . rrrn wrmrn. v rrn mn.. n nm. n n n vnm r -m n-rn n -mmnr n m.rrm v rrvn mrm,nnmrnr rnrw-nn mrnn.. nv =nnm. r nr m w.. v rrn -rnm-nnm. m n r r wrr nnrmz mnnr. mnmn.r,nn,nnnnnrvnr.rm r-rmnmnr . rmnn nmn-nr r-rrn mmnn.. n =nm. m rrnn rmr v -. - x - rmmmrv nvn n-r m rn nn rmrr mnr wn-r -.n - n nm wn-r rrrnmnnrn - . v.r nnr -r m rn n. v n .-v. wvr nrmzn nvmx vrn nw rnv r v rnv .rnr n nn-m nmxn nv wr rrrv n rrrnmr -nr. mn - n-r n. n =n n rmx-. v vn nrnn rn-r vrrn.-. ωrm.m n =nn v. ωnn r mnnnv. vn v w ωrrrmm mrmn r-rxmn. wnrwr mxnnmn n r-r , r v. rrnnnnn r .- nvnrmmnnm ω w rm - n. v v nnn r . mmrrnnr- . rrnn nn r n rn ω nn n . rrn vr n = vnrn rzm r,. , n r zn n r, mn. vn v v r ω vr wrnrn r r n = nnnr rr m nrmz .n r wr rrn mn ω n =nn wn vr rzm. . rnnnn vnr mnrnr-mnmnr r .nnnn ω n n. m rrnn r , n.r,-znrrmnr,r,-nn vrrm rrzrm ω. nx vrrn v r rrnn v m <rn n. n =nnn r r . m rrnn .--. n r-r rrrn. n -n ω rnrm rnr-.n.r m nn mn n.,n m r n.mrrr nmrnrrrm rrn nn n r x nn , n nnn rn m r- n nn. -rnnn, n.n rn, vr n w n rn---rn-rrr r wvrmn-r mn nn,n,,-nr. wrwn w n,r mn , rrnnzm m nn. , -nnrmr rvr r n-nr rn ,nmr rnrn r, n r n n r,vr, m, vr r rrnrn , n m , zm. ,rn n , n rn , ,x n , n nm-vn,-r,rnnnr.nrnr,rmn ,xm xm:vmn nmxr n v n nn mmmn - 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'-, v - . -,.- =. .. , - - . , - - - . ,.– , , - - . :. - - ., , ., - - .. , > , . .-, . - - , .- . -, .- .- - ., .. . - . - ,, , –- . • • -, - . , .. , , - . . ,xm: -ruurnn wnmx rv’rn ω n - , .. = ., =.,. ., ., - . , --. , . - , v , -, - - . -, , .-. , - .. . ’- , - . -. , ,,– . - - , - .- , . , - xm: r . '- n nnnrnrmnxv vrnrrnv rnn nn rn, nrrrwnnrnn rvnr.r v. m, w -r rmnn '- n,. nmx..r r n - , w r rm nrn m ’- r rn r’- vr’- .. r rnrnn rn, .n -r m nn n n rnrr . n rrzn rn rv mn r . r nr w - n ’- . w m,nr mn- r n r. rrnnn, w nx w r r m n n mx. rrm - v vrnnnr nm mn n -. nn vrn wnv mr wnr nrrr nv -r nrnmvxn,x vrn w >- nnmnnv mr rrnv r rnrvrn ,x, vr,n, , .-.wr w - , wrrm - n n ..nrn, vrn r n vnrn-,v nwrrnr-n rxnr-n-. -vr nn -n.,. mrvr,x mnrnr n n vrn vrn rnn nn-nn nn r , wr r vrnx nv, vrnnn r’- r’- . , wnmx- r nx. .n.n nv.n ’- mr w nmx mr wr ..rr r w nn ’- ’- ,nnrmnmx- nrr r ,mn w r rxn’ n rrnrm’- ’- n v -- .. ,nmx-r n v r m n nvnmx vrnnmx- nrnnmx- , w nxn - xrmnn x n-nr ωn w.n,.. vrn, ω rn nn mmrr -r n rn. ω rn n w . ω,n-rn ,wvr, wnrmmrrx,n. r’- nr,n nnmxnr rn r rn rnn nv vrrnn. v xm:n vvnm n wnmxrmn vr - , - ,. , .. ..,. ’ − − -. , .. - - , .:-,. , .. - -. - : . - - - - ., -12.,.--., , v . , .. -1. ... .,. ..313. , . . , , 1 , . . .3.. -..13-. ,. . . . . . , .. ... - , - - .. , - . , , - - - - - , .-. -. , , . ’-. , ’- , - , , . , .. v . , r nnurnnn,.. wrxrrrnm vrn wr m- n mrr w mm . m xrn ω wr n rn nrr . r- n mrmnmnmnr wrr n nr n mn . nr nrm wr.rmr n rnr n . n mnrr wrr rmrnr n nrm v nrr rrrrnnnn. ωr-rn n-. .r wrrrnnnnn nrr n-rn r n w ω n rrnn ω nr rnr n n- nnr rvnn n n wr. m rrnn wrw wnn.. n xr m-rm n. r rn mn rr’r .  vm rmn r nmn wr nr nnrnr nn m n mn r wn mn rr’ n r m rxnrzr ,  °rr rm r nn vm r . r w nv n ,. vm r n rrmnm,nr.r vrm mn, w  mnn. mx w vrxr  nn w r n r w vm nr n,  . rnnmm ,n wr-r r  - n . n w rmv r,. m r m mn. w vn r m rmn nn.n n wrr n rr n nr rvr r nr n v -.rnrm wr n nnn r–rn r    mm, rm r r, mmn.rn n  °r , n wr wnuurn.m rn wr n nr. wrn mrr °n 2 m r. mnn - n ' -' m wmr mnn wn rm r n. r- m wrnr w vmvm'nmm n m . n nnrmnvv, vv vnrm m-r w n wr n rm rn. wr mnn nn r rnm r vnr. mnmnn. wr n r .4v rn n. rr- w wr n -w -m ω xr rnn,rn   r n w  n r,n  n mmnmn.  mn nvrnr w.n r, r n wn rrmv m,n wr nnr r r w, n nn mnnn.  rmnrxrmn w nrn nx r-w ω-mnmrr . wrrv r rn -,.-,.,. n.n - ,r-m rr ’- - . nnm ,mx w-r r nn’-n n n  r r: mr   n r – , rn, m , n wnrrrm:n n. w rrmn rm r- n vm  °, mn °, mn °,. w r-m  rrnn nrn-rn wrrr n . r v m wrn n rrn.rrnrn rr, nn n rn n - ,n m vr x rrr m n r n n .6mr.rnnn wrmrr n m mm r m n mn nvrn r r . r, rmxn. m - mm.rn- n mx wrn r mn n rw r vr rn nn, nrwnx nn n nmnr- - n w rn...nn--rn, -rr rv rrnmnn-wn-mn rn ,,nrnn.- wrrrn . w wr 6 w mmnr wr >m, mn mn , rmxn. m -6.rr w mmrn wrrnnmxn mmr,r r . n -ruu unn m n unn.mxn. m - . r nn w rrm rv rrr.nnr, r - mrrn rmx,rm w r r rrm m nm nr w xnn-nmn  rwrn rrvxr wr n.rrn rn ,nrn. n mn rr’ n n w n n .nvrnnrmn.rr wr nz n mrvn mn, ,r-n.mn n wr -ruu rnr.r ‘-v w’rrnrn . n . rr , – .nr,nm w rnnn r-- v w-r- n nnrm nxr n r r. - r- ω wr z nnrnrrnn n r n. r rr w rnn vrrrnrnn r n . n . ω . n nrmzrr wrnn - -wx mxr mn °. mxr. ω, n r n wr wn r rr. nn r n w . n rrrnnm mrv , rn,nm mn-n .rrm,wn, nrmrrn wrnnrvn vrn n.wmr wnrr n n n - rrz v mn.n w : .wnnn r wr:rrn nrn m numn-nw,. un-.nn5n u-rnr n n r, wr r nn n n rn,nn,mrxwxzr,mnrnnzmmm, n– n nnmxr-v xr mn w n n vrnxr nm, n rmn, n n m - ω n m,n - rn xn , wn rn wrrn .. nn ,– .r nvnr,n rrmn nz- rn wrrrmn r n - rn mn rr’r .rmnnr w n n n nnm w r rnrn r wrrv ∼ -rn n m. - r n rn nrrnn rr nr nr n n,wnnm r n,nrn nnrn.n rmn n-r wr nrm r mn nn.n rnn, wnm m w r nvr z nvr , nnrm n rr’r .m rr wr n nmn .. n. wrnz n n-r - wrvrn nm r wr m mn rrn nm.nn n mm rnnnrnr nn ,-r wr rn m zn n -r wrrrnn nn rnnr. n nn mn vn r vr- nnnrr. - -nn.n w- r rrmrvmrmn nrr: rnn . rnn r, nmr-m , - v ...m w mnr m n rmn . r r, : n .x.r.™ .rr,nm wrmn nzm . rnr n xrrr™, nrnvrrmnn r n,w rn zn r mr r wrr,rrmn wnnn: °r, ° w r , r r nr ° n r n x °rx.rrmn,nr. r mn rr nn n m rr wr n zn n n nv rm w nrn. r- vrurrnun.nrrr r wrr r r. rvm wr r .r, vr wrrnr n nmu. xrmn mn n . rmrnvrn. nm wrw, n wrnrrmn rn wr rv rmn n. . wr nnn rnn-rrmr, n rm nx nvr nm nm- rn nmrr- rr wnr rm n.°, w .nr- r n nr r w rmnrr vnv rrvn. wr r– nrrrvnrurn n.wrn m rrn.nrrrvnrr12 1w3rn w vr v n rn nrnn ,x v rx v .nm wrn z m rmn wrmnr w m mmnn 2n n rx . rn w mmnn nnr mrr.n w mnnn n mnrn mn,2 mnrn n. nn wr mm n r rn mnm , r n r n m r, r v. n mnn. m wrmnr rnrn,. m w, rn mmrnrnrr,,w n w rnn n w mmnn rnrr n mnnn w.-. rn .x13vm w .rn nn rx, x vrr: rr wrn mrrnn:nrn n r m r rx . mmrvnr . mmnr rr . mm m r - mn. n. wnwn r wmrrrm n mm n r nr- v mn rn n,n wn w rn r n. mm nn.nnmn wrrrn mnzrx.rn2rnn wnrnrrn wrn rn n. v . w n rn rr mr .-, m ,. m ,nvnrn °r mnn. nn n °r, wn rnm’rn n. wr nz n 2nmm r w vn nnnnrm r n . m v nrr vn v r n rnn r vn.rn w ,nvr w z rnn vr, m wrr w vrm r w vr . nn–rmn rm r n, mn mmnw- r mn.vr r r mn wr n n mmw- n x wrmvvr nn–rmn rm r n rn mnnn w r n-,mnr.vr wnnn w n w wr wrrn mmrwn rnn w nrr. rmr r r mnm rrn rv nnvr mrr.. mmn nn r,nv wr.rmn n rrnnr m rn mn rr’ rn. rn w mr  nmn rn  nm r w m mrn wrn. r,r rn -n rn rm mr nv mrr rnrmr vn n mrn rnv wr rn’n rr nn-r. w-n nrnmn.rn wr nrrnw -n wrmn.-nn m. r n rm n rn mxrn rnxr n rn mn rr’nrn.r vr mr .n, m, w,r. n rmmnr n r n, mn wr n,r.m-mrx , - rm r.rnnnrn wr rmn n r nn m- rm n wn mn rr’r .nrnnm rn nrmz rn ww rnnnrnn m n wr vmvrrnn mrn: n rrnxmm n r - r : .rn wr rm m rr n - rrn n, mmnrnn wrrnnrr w m.mn n-r r mmnrnnm-rm.rnr :,n. n.. ,-.,nmn-rn, r nn r nn-:mn’ :. n.. n--. v n ,nm.-n wr xmnrm.nnnnn- wn nrm ™ vrn.- . r rmn n u xx nnmnmmmnnn-n-xn.rmm wrnz nnrn. wr n rn mn rr’ nz n x wmrnn mn n n unvn,mnr wnw rr nrnn.. xrn w rrm w rn mn w r vrrnr rm x vrrrn’nrn.rmrx , n.:m- nn w n wr mrrmrvn m .rn w m.nv vn mxr n w rmn w n. m nr wr nrn n rn r-m rrnrnn 2rr n . r v m v n.rnrn rr, r- “r rrnn wrn n r n r ,nn mvnrn xnrrrrn mmr. -rnn n, wnrwnn.nx nn nrn, nmn-rrr- r vr n n r.. -nmxn wr v - n n ,,n.rr w m rmn n m r6mr wrrnn-w - mn , rmrnn w. w n m n mn n rwxn. mr m-m..rn n mx wrn r rrn rv-rn..: urn ,– . r,:,.– mn . m -- - n . n, ,– rrnn rn.nn–n mn n rn nrnrr vn v rnnn: r nn rwn rnn wnnnrr.n, nnn wn MIRNKAFVEr VRLIYDv PNN. AA:TELI1N:RTTLnGSBARFDVeYNnHoFcLoAcRcRuIsAAYrKaESGoKGuLrTaYnGsT: KFALNSLKNLETAYKNFFRTVKSGKKVGFPRFRKKRTGE SSELTLLKAEETSWLSEVD GKILNVTVRRIHEGHYEASVLCEVEIPYL SYRTFTNNNIIGEGRLKLPKLGWVKTKGDI GTLSRRKKGSARYGKAKTKLARIHKRIVNPKARAPDKFFLAHAKGLVTDTVSGLIVKRDEFYAEIIVITGDTGEVHRLFKKPHDENMNRPKKNYRYRRLSATLLSKIRSLDRAKGAW AEGFIISRDTLLENYAKHAGAGWYYVGRRPLAVSSAKGVNSGPLYRFSPESNSHALTCLHVDVCGFKNPEVKNLAVRTWTCPNCGETHDRDENAALNIRREALVA PKKKRKE ID N:2:V4 NL V4: E IDN: : Bartte NLartca: E ID N: : nu cearocazaton E AFGAKLILNNSVLTKVNRLREITHAEYGKHNYFEFARSTVVLKCESVGEKIKPVYGLFPPARAFPRKKFKARATGGVEDSVYGRITKDFFTANNNIIGEGRLKLPKLGWVKTKGDI TLSRRKKGSARYGKAKTKLARIHKRIVNKRDFL IVTDGVRFKHENPKYYRSTLKRLRKA GEFIRDTLLENYAKHAGAGWYYVGRRPLAVSSAK HKLTTSLVREYEIIGTEHLKPDNMRKNRRLALSISDAGW AGIS GVNSGPLYRFSPESNSHALTCLHVDVCSGRFAKDNPPKEKVKKRNKLGAGVGRGTGWSTGCKPRNTCAGDEGTSHEDFREDPEKNKAKARLKNVIRREALVA SGGSSGGSESGSIEDTPNGT:SESATPESSxGGSS-GXGTSEN-x nerartca: v RMLISDEAVTELFE YSVHTEI FYD EWPMNRH:ALTLAKRATRaDEAREEVP:VGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRGGLVMNY YRMPRVFNAKKCAVMSCSAIGNAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDF M E IDN:1N-ABEartca: ISDEAVTELFYSVHTEFYEWPMCRVHMACLATGLAAMKIRHASRDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRGGLVMNYRL MPRVFNAKKASSINSGGSSRGIGGSRSVGVSFEGTVPRGNTSSKERSGAATAPGESSLSMGNGVSLSNYPGMNHRVEITEGILADECAALLCDFYR ARFVYNHFLARRIAAYKESGKGLTYGTSSELTLLKAEETSWLSEGVGDSKIFRANLKANFSVLVKRNLLYEPTNAAYAKNTFEFLRITNVRKTLSGGSK TTCCAGTTEAAGIGDAGNAG:GAA1T2ωRNA-eno-uman-1artca: GAACGTCTEGTCICDTTNAC:CCC1TωRNA-eno-uman-2artca: AGAACACCECATIADAGNAA:CAA1C4ωRNA-eno-uman- artca: AACTTTAGEATGITDCTNGT:TTC1CωRNA-eno-uman-4artca: GGCAGGACETGCICDTCNGG:ACA1GωRNA-eno-uman- artca: GCATTTCTEGTTITDTANAT:TTA1T7ωRNA-eno-uman- artca: v ATTATTGAETGTITDGANTC:ATA1AωRNA-eno-uman-7artca: CATAACAGECTAIADCANGT:TTT1TωRNA-eno-uman- artca: ATTGATGGECTAICDATNAT:CAG2AωRNA-eno-uman- artca: CTTAGTGAETGAIADATNAT:TTC2T1ωRNA-eno-uman-1 artca: GTGATGGGEAGCICDCTNTC:TTC2T2ωRNA-eno-uman-EMX1artca: GATTCAAGEAATICDCCNGA:AGT2GAωRNA-mn 117artca: GAGACGAGAACGCTGCGCTGAAAGCAAATTTCCTGTGGCCGCTGGTACACGGCTGATCTAGTGGGACTTGCCCGAACTGTGGGGAAACCCATGACC GTCAGTGTEGATIADGGNAT:CCG2T4taret1romExamesartca: GTGATGGGEAGCICDCTNTC:TTC2Ttaret2romExamesartca: GGCCGGCAETGGITDCCNCA:GCC2TCCTHCDGVCTrGGCoGzCCmGeGCRTNGAGGCAACATGCTTCGGCATGGCGAATGGGACTTTTTT

Claims

vm. n vrn mrn,rnn, - vn n rm rrnrvnn vnn n,r rvn n, mr r n :,rnvrn n, vnrnu r urn v :,n rrzn vn mnn,, rvn rnn:,, n n,n nn, - n u rnr rnn, n r urn r n : , n n rn nznr mzmnrnn,r nrr znnrnrm nn rn nnrr rm n r zn m . n mn vrnnr. rn m, wrn n r zn . nm vrrnn : . n r zn m mrrn nrn r n:m., wrn . nnr vrznn rn nn r n m, wrn . n vrn m rn:. m n r n r n m, wrn m n .m r , n , mrr r wrn .nn vr vnrn nrnmrn,nrn r n m, wrn . n , :. - wrnn vrnn mrn vrn rnmrnn n r nvnm m n,, -n n mn.v. m n mn n rn m, wrn n vn. mn n n , r nnn mrnnv ,rmrr w, w: rnr.n nnn mn . nrn r:n. mn nn m ,nn n . n nnnrnmn vr:nnr :. r n r n nn m , wr n nn m ,r r .nn n n nmzrrnrnnmn . nrxrnn vrmrn n n rn m,nr mmr r n mmmn,r rnrnr . xrnnn vr nr. xrrn mnr,nrrrmrrmnrnrxr n . m vmrmmnr,nr nr wrnnrmrrnn m,rrmn xr.r n . v mr r mnrnrn nnm nnrn r. , n n mnn,,n mx mrn vr r n r n m, nn nrrn. vrr

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