Waa maxay tallaalka mRNA
Tallaalka mRNA wuxuu u gudbiyaa RNA unugyada jidhka si uu u muujiyo oo uu u soo saaro antigens borotiinka ka dib isbeddelada ku habboon ee vitro, taas oo u horseedaysa jirku inuu soo saaro jawaab celin difaac ah oo ka dhan ah antigen-ka, taas oo kordhinaysa awoodda difaaca jirka[1,3].
Jaantus 1: Jaantuska jaantuska saamaynta tooska ah ee tallaalka mRNA [2]
Kala soocida tallaalada mRNA
Tallaalka mRNA waxa loo qaybiyaa laba nooc:aan soo noqnoqonmRNA iyois-weyneynmRNA: is-weynaynta mRNA kaliya kuma koobna antigen-ka bartilmaameedka ah, laakiin sidoo kale waxay dejisaa soo-celinta taas oo awood u siinaysa kordhinta RNA-ga gudaha iyo habka muujinta borotiinka.Tallaalada mRNA ee aan soo noqnoqoneynin oo keliya waxay dejiyaan antigen-yada bartilmaameedka ah waxayna ka kooban yihiin 5'iyo 3' gobollo aan la turjumay (UTR).Waxay bixiyaan dhiirigelin dhamaystiran oo la qabsi ah iyo difaac la dhasho, kuwaas oo ah situ antigen muujinta iyo gudbinta calaamada khatarta ah, waxayna leeyihiin codsiyada soo socda Tilmaamaha[2,3]
●Waxay ku siin kartaa dhiirigelin dhamaystiran oo la qabsi ah iyo difaac la dhasho, kuwaas oo ah muujinta situ antigen iyo gudbinta calaamadaha khatarta ah.
●Waxay dhalin kartaa jawaab celin difaac "dheeman", oo ay ku jiraan maaweelo kaftan ah iyo kuwa gacanta iyo xusuusta difaaca
●Waxay isku dari kartaa antigens kala duwan iyada oo aan la kordhin kakanaanta diyaarinta tallaalka
●Horumarka joogtada ah ee awoodda difaaca waxaa lagu gaari karaa tallaal soo noqnoqda, mana jirto ama wax yar oo ka jawaab celin ah oo ku wajahan sidaha
●Tallaallada mRNA-ga ee kuleylka leh waxay fududayn karaan gaadiidka iyo kaydinta tallaallada
Jaantus 2: Jaantuska jaantuska ee tallaalka mRNA iyo habka muujinta antigen-ka [4]
Tilmaamaha tallaalada mRNA
Marka la barbar dhigo tallaalada dhaqameed, tallaalada mRNA waxay leeyihiin habab wax soo saar fudud, xawaare korriin degdeg ah, looma baahna dhaqanka unugyada, iyo qiimo jaban.Marka la barbardhigo tallaalada DNA-da, tallaalada mRNA uma baahna inay galaan xudunta mana jirto khatar ah in la dhexgalo genome-ga martida loo yahay.Nolosha nuska waxa lagu hagaajin karaa wax ka beddel.
Shaxda 1: Faa'iidooyinka iyo khasaaraha tallaallada mRNA
|
| Faa'iidada | Nusqaan |
| tallaalka mRNA | Cilmi-baaris degdeg ah iyo horumarin, soo-saarka tallaalku wuxuu qaataa 40 maalmood oo keliya | Kicin jawaab celin difaac oo aan loo baahnayn
|
| Degenaansho la'aanta mRNA ee hoos timaada xaaladaha jireed, fududahay in la dhimo | Ma dhex geli doono genome-ka si looga fogaado isbeddellada daweynta ee suurtogalka ah
| |
| Looma baahna calaamad meelaynta nukliyeerka, qoraal | Waxtarka nukliyeerka badbaadada ayaa weli ah in la xaqiijiyo
|
Jaantus 3: Jaantuska socodka ee soosaarka iyo diyaarinta tallaalka mRNA [4]
Qalabka go'doominta ee fayraska RNA
RT-qPCR Fudud (Hal Tallaabo)
Xeeladaha la hagaajiyay ee diyaarinta tallaallada mRNA
Sababo la xiriira xasilloonida liidata ee mRNA lafteeda, hoos u dhac sahlan oo ay sameeyaan nukliyeerka unugyada, waxtarka gelitaanka unugga oo hooseeya, iyo hufnaanta turjumaada oo hooseeya, cilladahan ayaa xaddidaya adeegsiga tallaallada mRNA.Hufnaanta turjumaada waxay sidoo kale ciyaartaa door aad u xasaasi ah.Baabuurta geyntu waxa loo qaybin karaa fayraska fayraska iyo kuwa aan fayraska lahayn (oo ay ku jiraan liposomes, non-liposomes, viruses, nanoparticles, iwm.).Sidaa darteed, tallaabooyinka hagaajinta ee khuseeya ayaa loo baahan yahay.Kuwa soo socdaa waa istaraatijiyad horumarin farmaajo ah oo loogu talagalay diyaarinta mRNA[2]
1 Isku-dubarid koofiyadaha analoogga ah ama isticmaal ensaymes capping si aad u xasiliso mRNA una kordhiso tarjumaada borotiinka adigoo ku xidhaya qodobka bilowga turjumaada eukaryotic 4E (EIF4E)
2 Hagaajin walxaha ku jira 5′-aan latarjumin gobolka (UTR) iyo 3′-UTR si aad u xasiliso mRNA una kordhiso tarjumaada borotiinka
3 Ku darida Poly(A) dabada waxay xasilin kartaa mRNA waxayna kordhin kartaa turjumaada borotiinka
4 Nucleosides wax laga beddelay si loo yareeyo firfircoonida difaaca jirka oo loo kordhiyo tarjumaada
5 Daawaynta RNase III iyo nadiifinta dareeraha borotiinka degdega ah (FPLC) waxay yarayn kartaa firfircoonida difaaca waxayna kordhisaa tarjumaada
6 Kor u qaad taxanaha ama codons si aad u kordhiso tarjumaada
7 Isu keenida arrimaha bilowga tarjumaada iyo habab kale oo lagu beddelo tarjumaada iyo difaaca jirka
Jaantus 4: In vitro transcription (IVT) mRNA wax soo saarka iyo habka isu-ururinta [5]
Diyaarinta baaxad weyn ee DNA plasmid
Nadiifinta DNA-da Plasmid waxay inta badan ka saartaa wasakhda sida RNA, endotoxin DNA-da goobabada furan, borotiinka martida loo yahay iyo aashitada nucleic-ka ah ee martida loo yahay, waxayna caadi ahaan u beddeshaa plasmid-ka dib-u-habaynta E. koli.E. coli waxa uu maraa halsano cufnaanta sare leh, ka dib kala soocida dareeraha adag, iyo ururinta E. coli.E. coli-ka ayaa markaa lagu sammeeyaa lysis alkaline, kala soocida dareeraha adag ee centrifugal iyo caddaynta microfiltration ka dib lysis, ultrafiltration iyo xoojinta ka dib caddaynta, ka dibna nadiifinta chromatographic.
Nadiifinta DNA-da plasmid:
Qalabka yar yar ee Plasmid General
【1】苗鹤凡, 郭勇, 江新香.mRNA疫苗研究进展及挑战[J].免疫学杂志, 2016 (05): 446-449.
【2】Pardi N, Hogan MJ, Porter FW, iyo al.tallaalada mRNA - waa cusub ee tallaalka[J].Dib u eegisyada Dabeecadda Helitaanka Dawada, 2018.
【3】Kramps T., Elbers K. (2017) Hordhac Tallaalka RNA.Gudaha: Kramps T., Elbers K. (eds) Tallaalka RNA.Hababka in Molecular Biology, vol 1499. Humana Press, New York, NY.
【4】Maruggi G, Zhang C, Li J, iyo al.mRNA oo ah Farsamo Isbadal ah oo loogu talagalay Horumarinta Tallaalka si loo Xakameeyo Cudurada Infekshinka ah[J].Daaweynta Molecular, 2019.
【5】Sergio Linares-Fernández, Céline Lacroix, ,Daawaynta tallaalka mRNA si loo dheelitiro jawaab celinta difaaca jirka, Isbeddellada Daawada Molecular, Cadadka 26, Issue 3,2020, Bogagga 311-323.
Waqtiga boostada: Agoosto-05-2021
