(2010) The effect of morpholino knockdown of genes TFEB and PKD2 on gene CDH-17 and green fluroscent...

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Transcript of (2010) The effect of morpholino knockdown of genes TFEB and PKD2 on gene CDH-17 and green fluroscent...

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    [bkkbdl 2

    Klfifc|

    Wea annaow fn jf{xefildf mdfomgfd fn cadas WNAK bdg XMG; fd aux{asslfd fncada OGE23 bdg c{aad niq{fsoadw x{fwald %CNX, lweld wea x{fdaxe{fs fn Gbdlf

    [a{lf%pak{bnlse,

    Wbesaad [bkkbdl_asslfd dqjka{0 8858>*87>

    Ead{lof Elce _oeffi

    Ofqdw{|0 Qdlwag _wbwas

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    _qxa{~lsf{0 Oeb{ias Nbiis

    Sf{g Ofqdw0 ?)

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    [bkkbdl ;

    Bksw{bow

    Wea auxa{ljadwa{ ld~aswlcbwag wea mdfomgfd %oassbwlfd fn x{fwald x{fgqowlfd, fn wf cadas)

    WNAK bdg XMG;) bdg wea sqksarqadw annaow qxfd wea aux{asslfd cada OGE23) bdg c{aad niqf{asoadw

    x{fwald %CNX, lweld wea x{fdaxe{fs %nl{sw swbca mlgda|, fnGbdlf [a{lf %pak{bnlse,$ Ljxilobwlfds fn

    {asqiws fqig ka ~lwbi ld bssassldc wea {asxaowl~a cadas ljxf{wbdoa ld wea x{fdaxe{lo cada {acqibwf{|

    ela{b{oe|$ Kfwe cadas a{a mdfomag gfd %oassbwlfd fn x{fwald x{fgqows, qsldc jf{xefildfs %j[DB

    ldelklwf{s,) bdg aux{asslfd fn CNX bdg OGE23 a{a jabsq{ag {asxaowl~ai| qsldc wf glnna{adw ildas fn

    pak{bnlse bdg ofjxb{ag bcbldsw bd ldgaxadgadwi| o{abwag rqbilwbwl~a sobia$ B w{bdscadlo ilda fn pak{bnlse

    bs qsag wf swqg| CNX aux{asslfd) biifldc nf{ niqf{asoadwc{aad ~lsqbilpbwlfd lweld wea x{fdaxe{fs)

    bdg wea saofdg ilda a{a lig w|xa BK pak{bnlse) qsag wf swqg| OGE23 aux{asslfd k| swbldldc fn OGE23

    j[DB$ Bii ? cadas swqglag bnnaow x{fdaxe{lo ga~aifxjadw bdg weqs lw bs e|xfweaslpag webw mdfomgfd

    fn XMG; bdg WNAK fqig {asqiw ld ldelklwag CNX aux{asslfd lweld wea x{fdaxe{fs) gqa wf gbjbca) bdg

    iassa{ aux{asslfd fn OGE23) gqa wf wea e|xfweawlobi gaxadgado| fn OGE23 aux{asslfd qxfd fwea{ cadas

    ld~fi~ag lwe x{fdaxe{lo ga~aifxjadw$

    Wea {asqiws ldglobwag b ibom fn slcdlnlobdw glnna{adoa ld CNX aux{asslfd bdg OGE23 aux{asslfd

    kawaad wea auxa{ljadwbi %JF ld`aowag pak{bnlse, c{fqxs bdg ofdw{fi c{fqx %qdld`aowag pak{bnlse,$ Fda

    {absfd nf{ WNAK JFld`aowag c{fqx dfw glsxib|ldc bd| glnna{adoa ld aux{asslfd ofqig ka wea aulswadoa fn

    bd lsfnf{jlo WNAK x{fwald lwe b w{bdsibwlfdbi swb{w glnna{adw n{fj ebw wea jf{xefildf bs bkia wf

    kifom= jabdldc WNAK jlcew dfw eb~a kaad nqii| mdfomag gfd$ Bs nf{ XMG;) xa{ebxs mdfomgfd fn

    OGE23 bnnaows aux{asslfd fn XMG; {bwea{ webd ~loa ~a{sb) elia CNX aux{asslfd bs dfw slcdlnlobdwi|

    glnna{adw kaobqsa XMG; mdfomgfd bs dfw adfqce wf biwa{ niqf{asoadoa$ Nf{ bii nqwq{a {axawlwlfds)

    glnna{adw ofjkldbwlfds fn mdfomgfd bjfdcsw wea ? cadas fqig ka {aofjjadgag wf xfsslki| |laig

    slcdlnlobdw {asqiws$

    ]Sf{g Ofqdw0 ?88 f{gsZ

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    Wbkia fn Ofdwadws

    _aowlfd Xc$

    Bksw{bow$$$;

    Ldw{fgqowlfd$$$$$7 sqkqdlws, jf{xefilda {ldcs

    ofddaowag k| xefsxefglbjlgbwa c{fqxs bs b kbomkfda %Dqssiald^fieb{g) bdg Gbej

    27;,$ Wf qdga{swbdg ef wea| nqdowlfd fda jqsw ofdslga{ wea oadw{bi gfcjb fn

    klfifc|) eloe swbwas webw GDB jbmas [DB jbmas x{fwald= wea s|dweasls fn wea

    x{fwald kaclds lwe wea GDB fn b sxaolnlo cada kaldc w{bdso{lkag ldwf jassadca{

    [DB %wea {asqiwldc w{bdso{lxwls b saw fn ld sw{qowlfds nf{ wea bssajki| fn wea

    x{fwald, %[qssaii 55,$ Wea j[DBs sarqadoa ls wead qsag wf bssajkia b x{fwald

    lweld wea {lkfsfjas %w{bdsibwlfd,%[qssaii 22,$ Wea j[DB lws ain ls sldcia sw{bdgag)

    sf wea xq{xfsa fn b jf{xefildf ls wf kldg wf wea j[DB dab{ wea swb{w fn ea{a ls lw

    wf ka w{bdsibwag %t;1 kbsas,) wf x{a~adw bssajki| fn wea x {fwald wea cada ls ofgldc

    nf{ %Klii) Xawpfig) Oib{m) _oeljjadwl) bdg Amma{ >

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    [bkkbdl 5

    x{fwalds sqoe bs WNA?&WNAO bdg kldgs wf akfuas %saowlfds fn b cada ifobwag `qsw

    bkf~a wea x{fjfwa{= wea ifobwlfd ea{a w{bdso{lxwlfd nf{ wea cada kaclds, lwe wea

    sarqadoa OB\CWC %+Eqjbd Cada Ofjxadglqj+,$ Wf ebw annaow wels kldgldc ebs

    fd cada {acqibwlfd ls dfw nqii| qdga{swffg$ Ld jloa ajk{|fs) jqwbwlfd fn WNAK

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    {asqiws ld wea gabwe fn jloa lweld 22 gb|s) kanf{a f{cbds a~ad ga~aifx

    %_waldc{ljssfd) Wassb{fiif) [alg) @admlds) bdg Dabi 7>83,$ Ld eqjbds) bd f~a{x{fgqowlfd fn

    WNAK x{fwald ls bssfolbwag lwe {adbi sb{ofjb %jbilcdbdw wqjf{s fn wea mlgda|, )

    bdg ld ab{i| ga~aifxjadwbi swbcas fn jbjjbilb) WNAK aux{asslfd obd ka sa ad ld wea

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    Wea annaow fn WNAK mdfomgfd fqig ka ljxf{wbdw wf swqg| ld {aibwlfd wf oge 23

    aux{asslfd) cl~ad wea bomdfiagcajadw WNAK xib|s b {fia lweld wea mlgda|) kqw

    wea ibom fn ldnf{jbwlfd bs wf ebw {fia lw aubowi| xib|s$

    Wea saofdg cada mdfomag gfd ld wels auxa{ljadw bs XMG;) b cada eloe

    ofgas nf{ xfi|o|swld; x{fwald) eloe ld ofd`qdowlfd lwe xfi|o|swld2 x{fwald

    %ofgag k| cada XMG2,) x{fjfwas glnna{adwlbwlfd) c{fwe bdg ga~aifxjadw fn oaiis

    lweld wea mlgda|s) ldoiqgldc olilb ifobwag fd wea mlgda| wqkqias %Cadawlos Efja

    [ana{adoa,$ 31 jqwbwlfds fn wea XMG; cada b{a bssfolbwag lwe wea xfi|o|swlo

    mlgda| glsabsa= {asqiwldc ld wea c{fwe fn o|sws fd wea mlgda| %Cadawlos Efja

    [ana{adoa,$ Mdfomldc gfd XMG; bdg sw qg|ldc wea ljxilolw annaows qxfd OGE23

    aux{asslfd fqig ka ljxf{wbdw ld aswbkilseldc lws ljxf{wbdw lweld wea x{fdaxe{lo

    cada {acqibwf{| ela{b{oe|$

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    Gaoef{lfdbwa ajk{|fs n{fj aboe glse bdg qsldc x2888 lwe oqw wlx$ Bgg aboe oiqwoeld saxb{bwa Axxadgf{n wqkas$ Bsxl{bwa bd| ianw f~a{ sfiqwlfd$

    Ajk{|fs jqsw ka xqw we{fqce b {ae|g{bwlfd sa{las02 bse lwe >>! JaFE %jawebdfi, & ??! 2uXKW %XK_ lwe Waad;8 bggag,2 bse lwe ??! JaFE& >>! 2uXKW7 bseas lwe 288! 2uXKW

    2 ji wfwbi nf{ a~a{| bse lweld wels x{fwfofi) 128 jldqwas) fd c|{bwf{| sebma{ %;8

    ?8 {xj,$ Bsxl{bwa sfiqwlfd kawaad a~a{| bse$

    X{ae|k{lglpa ajk{|fs ld 188 qI 2uE|k jlu - w[DB bdg eaxb{ld %E|k-,) nf{ 42efq{ bw >1 O$

    Gliqwa x{fka swfom bw b 20288 wf 20?88 gliqwlfd ld 2uE|k-) x{aeabw bw >1 O) wfwbisfiqwlfd obd ka 188 qI nf{ 2818 ajk{|fs lweld fda wqka$ Auxa{ljadwa{ qsag

    bdwlsadsa glcfulcadldibkaiag {lkfx{fkas gaslcdag wf gawaow oge23 j[DB$

    Bsxl{bwa x{ae|k{lglpbwlfd sfiqwlfd n{fj ajk{|fs) bdg {axiboag lwe wea x{ab{jag2uE|k- ofdwbldldc gliqwag x{fka$ Xiboa wqkas kbom ld >1 O ad~l{fdjadw f~a{dlcew$

    Gb| ;

    Sb{j 2uE|k jlu lwefqw bggldc w[DB bdg eaxb{ld %E|k, bw >1 O$

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    Ajk{|fs jqsw ka xqw we{fqce wea nfiifldc bse sa{las ld b >1 O bwa{ kbwe2 rqlom bse lwe x{ab{jag E|k2 bse lwe >>! E|k & ??! ;u__O %sbildasfglqj olw{bwa kqnna{, nf{ 28 jldqwas2 bse lwe ??! E|k& >>! ;u__O nf{ 28 jldqwas2 bse lwe ;u__O nf{ 28 jldqwas; bseas lwe ;u__O nf{ ;8 jldqwas

    Nfiifldc bse sa{las gfda fd c|{bwf{| sebma{ %;8?8 {xj,

    2 bse lwe >>! 8$;u__O& ??! 2uXKW nf{ 1 jldqwas2 bse lwe 8$;u__O& >>! 2uXKW nf{ 1 jldqwas2 bse lwe 2uXKW nf{ 1 jldqwas

    Bgg kifomldc sfiqwlfd ofdslswldc fn 2uXKW - ; jc&ji K_B - ;! cfbw sa{qj$ 2 jIsefqig sqnnloa$ Kifomldc sfiqwlfd ls gaslcdag wf kldg wf bd| x{fwald slwas eloe gf dfw

    eb~a x{fkas bwwboeag) {agqoldc kbomc{fqdg swbldldc$

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    [bkkbdl 23

    Gliqwa b sjbii ~fiqja fn bdwlGlc BX %bimbilda xefsxebwbsa, ld kifom sfiqwlfd= 2028$Wead bgg 2 qI fn gliqwag bdwlGlc BX wf

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    [bkkbdl 25

    [asqiws . Gbwb

    Wbkia 7 Wea nfiifldc wbkia glsxib|s wea xa{oadwbca fn nqii aux{asslfd fn CNX bdg OGE23j[DB bjfdcsw ajk{|fs boof{gldc wf eloe cada bs mdfomag gfd$ Wnak aux{asslfd*

    Wea annaow fn jf{xefildf mdfomgfd fn cadas WNAK bdg XMG; fd Obgea{ld23 bdg c{aadniq{fsoadw x{fwald %CNX, aux{asslfd ld wea ga~aifxldc mlgda|s fnGbdlf [a{lf %pak{bnlse,0

    Xa{oadwbca fn nqii aux{asslfd fn CNX bdg OGE23

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    [bkkbdl ;8

    WNAK %288! nqii aux{asslfd,) sf xa{ebxs lw ls qdga{swbdgbkia webw lw fqig dfw iljlw wea

    aux{asslfd fn bdfwea{ cada sqoe bs OGE23$ Efa~a{) wf nqii| qdga{swbdg wea annaows fn XMG;

    fd WNAK bdg OGE23) fwea{ jawefgs lii daag wf ka qsag) sqoe bs ldo{absldc wea aux{asslfd fn

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    aux{asslfd fn XMG; bdg gfoqjadwldc eawea{ f{ dfw lw ldo{absas&{agqoas aux{asslfd fn wea wf

    fwea{ cadas$ Wels jawefg fqig eaix ~a{ln| wea x{fdaxe{lo cada {acqibwf{| ela{b{oe|) {bwea{ webd

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    WNAKs {fia ld wea x{fdaxe{fs fn pak{bnlse ls swlii dfw nqii| qdga{swffg) eloe ls e|

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    jawefgs) sqoe bs qx{acqibwldc ldswabg fn mdfomgfd) fqig eb~a wf ka ljxiajadwag wf

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    qdmdfd ea{a wels lsfnf{j ls aux{assag bdg wf ebw ofdoadw{bwlfd) kqw wels ibom fn nqii

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    fqig ka ld`aowag bifdc lwe wea f{lcldbi WNAK JF$

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    [bkkbdl ;2

    Nf{ nqwq{a {axawlwlfds fn wels auxa{ljadw) sfja oebdcas fqig adsq{a b lga{ {bdca fn

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    nf{ fdi| sf ifdc,%Klii aw bi$) >

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    [bkkbdl ;7

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  • 8/2/2019 (2010) The effect of morpholino knockdown of genes TFEB and PKD2 on gene CDH-17 and green fluroscent protei

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    elia wnak aux{asslfd ls jqoe k{fbga{$

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    Bgg ?8 jI fn [F %{a~a{sa fsjfsls, bwa{ wf b ;18 jI A{iadja|a{ nibsm$ Bggalcewag bcb{fsa wf bwa{ bdg sl{i wf jlu$

    Sbse bd| auoass xfga{ n{fj wea slgas fn wea nibsm qsldc [F bwa{ f{ gE;8%galfdlpag bwa{,$ Gf dfw qsa jf{a webd 2; jI$

    Xiboa nibsm ld jlo{fb~a nf{ ; jldqwas wf jaiw wea bcb{fsa$ _wfx a~a{| ?8 saofdgs wfwbma fqw nibsm bdg sl{i$ Qsa eabw {aslswbdw cif~a lwe ebdgildc wea efw nibsm$

    Bnwa{ bcb{fsa ls gfda jaiwldc) biif nibsm wf slw bdg offi gfd nf{ 28 jldqwas$

    Bgg ?8 jI fn jawe|iada kiqa %wf x{a~adw c{fwe fn bicba, wf wea sfiqwlfd ld weanibsm) bdg sl{i wf jlu$

    Xfq{ sfiqwlfd ldwf b 28 oj xaw{l glse$

    Ob{anqii| ib| gfd jlo{fld`aowlfd jfig fd wea sq{nboa fn wea sfiqwlfd swb{wldc n{fj b

    slga agca wf x{a~adw kqkkias nf{jldc ld wea w{fqces fn wea jfig$

    Biif 128 jldqwas nf{ sfilglnlobwlfd$

    Qsa sxbwqib f{ weld kibga wf x{| fqw jfig$ Xiboa glse ld b {an{lca{bwf{ nf{ 2; jldqwaswf adsq{a bcb{fsa ls sfilg$

    Nlcq{a ;

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    [bkkbdl ;3

    Bomdfiagcajadws

    Wea{a a{a jbd| ldgl~lgqbis) ef lwe weal{ iljlwiass xbwladoa bdg cqlgbdoa) jbga wels

    assb| xfsslkia wf ka {lwwad$ Efa~a{) wea kqim fn j| bxx{aolbwlfd ls asxaolbii| gqa wf G{$

    @bjas Ilswa{ fn ^l{cldlb Ofjjfdabiwe Qdl~a{slw| nf{ wea ofdoaxwlfd fn wea f{lcldbi x{f`aow

    %wea {fia fn wnak fd ajk{|fdlo ga~aifxjadw, bdg biifldc ja wf xa{nf{j {asab{oe lweld els ibk$G{$ Ilswa{ x{f~lgag ja lwe wea daoassb{| arqlxjadw= sqxa{~lslfd ead gabildc lwe bgqiw

    pak{bnlse bdg ebpb{gfqs oeajlobis= bdg w{bldldc ld elcea{ia~ai x{fwfofi daoassb{| nf{ wea

    auxa{ljadw$ Bii fn j| rqaswlfds a{a wef{fqcei| bdsa{ag k| G{$ Ilswa{) annaowl~ai| eaixldc jawf cbld b nqii c{bsx f~a{ a~ad wea jfsw waoedlobi b{abs fn j| {asab{oe$ L fqig bisf ilma wf webdm

    J{$ Oeb{ias Nbiis) j| sqxa{~lsf{ bdg xe|slos waboea{) bdg J{s$ X{lsoliib Klggia) wea LK

    off{gldbwf{ bdg j| weaf{| fn mdfiagca waboea{) nf{ x{f~lgldc lgabs nf{ {a~lslfd bdg glilcadwi|{abgldc we{fqce g{bnws fn wea assb|$