Abstract
Protein transduction using cell-penetrating peptides (CPPs) is useful for the delivery of large protein molecules, including some transcription factors. This method is safer than gene transfection methods with a viral vector because there is no risk of genomic integration of the exogenous DNA. Recently, this method was reported as a means for the induction of induced pluripotent stem (iPS) cells, directing the differentiation into specific cell types and supporting gene editing/correction. Furthermore, we developed a direct differentiation method to obtain a pancreatic lineage from mouse and human pluripotent stem cells via the protein transduction of three transcription factors, Pdx1, NeuroD, and MafA. Here, we discuss the possibility of using CPPs as a means of directing the differentiation of iPS cells and other stem cell technologies.
Highlights
The CPP sequence was originally found in natural proteins as the HIV trans-activator of transcription (TAT) [7,8] and the Drosophila melanogastor homeodomain transcription factor Antennapedia [9]
In contrast to directing the differentiation of stem cells, there is some difficulty in reprogramming somatic cells to InItnrdoduucecdtiopnluripotent stem (iPS) cells and the direct conversion of somatic cells to other cell types with protein transduction
Practical protocols are desired regarding protein-mediated direct conversion. It has been shown by many reports that some steps in iPS cell technologies can be done by protein transduction methods (Table 1)
Summary
1. InItnrdoduucecdtiopnluripotent stem (iPS) cells are generated from somatic cells and they have a capacity to difIfnedreuncteidatpe liunrtiopomteunlttipstlemce(lliPtyS)pecesl[ls1]a. rTehgeeunseeraotfediPfSrocmellstoemchantoiclocgeilelss aindretgheenyehraatviveeamcaepdaicciintye tinovdoilfvfeersenthtieatkeeiyntsotempusltoifplreepcerlolgtryapmesm[i1n]g. ,TghenueseedoiftiinPgS/coerllretecctihonno, loangdiesdiinffereregnentieartaiotinve(Fmigeudriecin1)e. iPnrvootelvinestrtahnesdkueyctisotnepvsiaofcerlelp-proengreatrmatminigngp, egpetnideesed(CitiPnPgs/)ciosraremctieotnh,odanfdordtihffeerdeenltiivaetrioynof(Fpigeputriede1s)., Prercooteminbitnraanntsdpurcottieoinnsv, iaancdelll-aprgeneemtraotliencgulpesep[t2i]d.eTsh(iCsPmPse)thisoda ims estahfoedr tfhoarnthgeendeelidveelriyveorfy pveipativdieras,l rveeccotomrbsinbaencatupsreottehienrse, aisndnolarrigsek mofoltehceulgesen[o2m]. icThinistemgreatthioond oisf seaxfoergetnhoaunsggeneneeds.elTivheerryefvoirae,vtihraisl vmeectthoorsdbehcaasustheethpeoressisibniloitryisktoofsuthbestgiteuntoemfiocrinvtiergursa-mtioendioafteedxoggeennoeudsegleivneersy. Tinhetrhefeorme,utlhtiissmteeptshoodf hreapsrtohgerpamosmsiibnigli,tygtoensuebsetditiutitnegf/ocrovrrireuctsi-omnedainatdeddgiefnfeerednetliivateiroynin(tFhiegumruelti1s).tepIns ofthreisproregvraiemwm, inwge, gsuenmemedariitzinegr/eccoernrtecretipoonratsndindtihffiesrfeineltdiatainodn (tFhiegufuretu1r)e. Ipnotshsiisbirleitvyieowf,uwtieliszuinmgmthairsizme erethcoendtirnepiPoSrtsceinll tthecishnfioelldogainesd. the future possibility of utilizing this method in iPS cell technologies. FFiigguurree 11..CPCP-PmPe-dmiaetdeidatpedrotpeirnotteriannstdruanctsidonuctteicohnnotelocghineos liongireespriongrraempmroignrga,mgmenienge,digtiennge/coerdreitcitniogn/, caonrdredcitfifoenr,enantidatdioinffeorfeniPtiSatcieolnlso. fCiPPPS-cmelelds.iaCtePdP-pmroetdeiiantetdrapnrsodtuecintiotrnanmsedtuhcotdiosnamreeuthsoeddsfoarrekueysesdtefpors kineyiPsStecpesllinteicPhSncoelollgtieecsh. nTohleogrieeps.roTghreamremprinoggraomf smominagtiocf cseolmlsaitsicincedlulsceisdinwdiuthceYdawmiathnaYkaam-4anfaacktao-r4s ffaucsteodrstfousCedPPtos.CGPePnse. Gceonrreeccotirornectoiof ndoisfedaissee-asspee-csipfieccimficumtautitoantioins ipsepreforfromrmededbbyytthhee CCRRIISSPPRR--CCaass99 ssyysstteemm wwiitthhCCPPPP-f-ufusseeddCCasa9s9enednodnounculecalesaes.eT. hTehdeifdfeifrfeenrteinattiioatnioonf ioPfS icPeSllscieslldsirisecdteirdewcteitdh CwPitPh-fCusPePdtfruasnesdcrtripantisocnrifpatcitoonrsf.aOctSoKrsM. O, OSKctM4,,SOocxt24,, KSolfx42, ,cK-Mlfy4,c;c-TMF,ytcr;aTnFsc,rtirpatnioscnrifpactitoonr. factor. Int. J. Mol. Sci. 2015, 16, page–page; doi:10.3390/ijms161125986 www.mdpi.com/journal/ijms
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