Urukingo rwa mRNA ni iki
Urukingo rwa mRNA rwimurira RNA mu ngirabuzimafatizo z'umubiri kugira ngo rugaragaze kandi rutange antigene za poroteyine nyuma yo guhindura ibintu muri vitro, bityo bigatuma umubiri utanga ubudahangarwa bw'umubiri urwanya antigen, bityo bikagura ubushobozi bw'umubiri.[1,3].
Igishushanyo 1: Igishushanyo mbonera cy’ingaruka zo guterwa mu buryo butaziguye urukingo rwa mRNA [2]
Gutondekanya inkingo za mRNA
inkingo za mRNA zigabanyijemo ubwoko bubiri:kudasabamRNA nakwiyubakamRNA: kwiyongerera imbaraga mRNA ntabwo igizwe gusa na antigen igenewe, ahubwo inashyiraho uburyo bwo kwigana butuma RNA yongerera imbaraga uburyo bwo kwerekana poroteyine.Urukingo rwa mRNA rudakubiyemo gusa antigene igamije kandi rurimo uturere 5'n'uturere 3 (UTR).Zitanga uburyo bwuzuye bwo guhuza n'imihindagurikire y’ubudahangarwa hamwe n’ubudahangarwa bw'umubiri, cyane cyane mu mvugo ya antigen no kwanduza ibimenyetso by’akaga, kandi bifite porogaramu zikurikira Ibiranga[2,3]
● Irashobora gutanga uburyo bwuzuye bwo guhuza n'imihindagurikire y'ikirere hamwe n'ubudahangarwa bw'umubiri, cyane cyane mu mvugo ya antigen no kwanduza ibimenyetso
● Irashobora gutera ubudahangarwa bw'umubiri "uringaniye", harimo gukora urwenya na selile hamwe no kwibuka
● Irashobora guhuza antigene zitandukanye utarinze kongera ubukana bwo gutanga urukingo
● Gukomeza kunoza ubushobozi bwubudahangarwa burashobora kugerwaho hifashishijwe urukingo rwinshi, kandi nta gisubizo gihari cyangwa gito kuri nyirubwite
Shyushya inkingo za mRNA zihamye zirashobora koroshya gutwara no kubika inkingo
Igishushanyo 2: Igishushanyo mbonera cyurukingo rwa mRNA hamwe nuburyo bwo kwerekana antigen [4]
Ibiranga inkingo za mRNA
Ugereranije ninkingo gakondo, urukingo rwa mRNA rufite uburyo bworoshye bwo gukora, umuvuduko witerambere ryihuse, ntukeneye umuco wutugari, nigiciro gito.Ugereranije ninkingo za ADN, urukingo rwa mRNA ntirukeneye kwinjira muri nucleus kandi nta ngaruka zo kwinjiza muri genome yakiriye.Igice cya kabiri cyubuzima gishobora guhinduka muguhindura.
Imbonerahamwe 1: Ibyiza nibibi byinkingo za mRNA
|
| Ibyiza | Ikibazo |
| urukingo rwa mRNA | Ubushakashatsi bwihuse niterambere, umusaruro winkingo ufata iminsi 40 gusa | Kurura igisubizo kidakenewe
|
| mRNA ihungabana mubihe bya physiologique, byoroshye gutesha agaciro | Ntabwo azinjira muri genome kugirango yirinde ihinduka ryimiti
| |
| Ntibikenewe ibimenyetso byose bya kirimbuzi, kwandukura | Ingaruka za kirimbuzi z'umutekano ziracyakomeza kugenzurwa
|
Igishushanyo 3: Imbonerahamwe yerekana umusaruro winkingo ya mRNA no kuyitegura [4]
Foregene Virus RNA Igikoresho cyo kwigunga
RT-qPCR Byoroshye (Intambwe imwe)
Kunoza ingamba zo gutegura inkingo za mRNA
Bitewe nuko mRNA ubwayo itajegajega, kwangirika byoroshye na nuclease mu ngingo, uburyo buke bwo kwinjira mu ngirabuzimafatizo, hamwe n’ubusobanuro buke bwo guhindura, izo nenge zigabanya ikoreshwa ry’inkingo za mRNA.Gukora neza mubuhinduzi nabyo bigira uruhare runini.Imodoka zitanga zishobora kugabanywamo ibice bya virusi na virusi zidafite virusi (harimo liposomes, non-liposomes, virusi, nanoparticles, nibindi).Niyo mpamvu, ingamba zifatika zo kunoza zirakenewe.Ibikurikira nuburyo bwo kunoza imiti yo gutegura mRNA[2]
1 Sintezeza igereranya rya cap cyangwa ukoreshe enzymes za capping kugirango uhagarike mRNA kandi wongere ibisobanuro bya poroteyine uhuza na eukaryotic yo gutangiza ibintu 4E (EIF4E)
2 Hindura ibintu biri mukarere ka 5′-kudasobanurwa (UTR) na 3′-UTR kugirango uhagarike mRNA no kongera protein
3 Ongeraho umurizo wa Poly (A) urashobora guhagarika mRNA no kongera protein
4 Nucleoside yahinduwe kugirango igabanye ubudahangarwa bw'umubiri no kongera ubusobanuro
5 Umuti hamwe na RNase III hamwe na protein yihuta ya chromatografiya (FPLC) irashobora kugabanya ibikorwa byumubiri no kongera ubusobanuro
6 Hindura urutonde cyangwa code kugirango wongere ibisobanuro
7 Gutanga hamwe ibintu byo gutangiza ibisobanuro hamwe nubundi buryo bwo guhindura ubusobanuro hamwe nubudahangarwa
Igishushanyo 4: Muri vitro transcription (IVT) mRNA yo gutunganya no guteranya [5]
Gutegura nini ya ADN ya plasmid
Isuku ya ADN ya plasima ikuraho cyane cyane umwanda nka RNA, uruziga ruzengurutse ADN endotoxine, poroteyine yakira na aside nucleic, kandi ubusanzwe ihindura plasmide ya recombinant ihinduka E. coli.E. coli ihura na fermentation yubucucike bwinshi, hanyuma igatandukana-amazi akomeye, hamwe no gukusanya E. coli.E. coli noneho ikorerwa alkaline lysis, centrifugal ikomeye-yamazi itandukanijwe hamwe na microfiltration isobanurwa nyuma ya lysis, ultrafiltration hamwe nibitekerezo nyuma yo gusobanurwa, hanyuma kwezwa kwa chromatografique.
Isuku rya ADN ya plasmid:
Foregene Rusange Plasmid Mini Kit
【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, n'abandi.inkingo za mRNA - ibihe bishya mu rukingo [J].Kamere Isubiramo Ibiyobyabwenge, 2018.
【3】Kramps T., Elbers K. (2017) Intangiriro ku Nkingo za RNA.Muri: Kramps T., Elbers K. (eds) Inkingo za RNA.Uburyo muri Biologiya ya Molecular, vol 1499. Itangazamakuru rya Humana, New York, NY.
【4】Maruggi G, Zhang C, Li J, n'abandi.mRNA nk'ikoranabuhanga rihindura iterambere ry'inkingo zo kurwanya indwara zanduza [J].Ubuvuzi bwa molecular, 2019.
【5】Sergio Linares-Fernández, Céline Lacroix ,, Ubudozi bwa mRNA Urukingo rwa Balance Innate / Adaptive Immune Igisubizo, Imigendekere yubuvuzi bwa molecular, Umubumbe wa 26, nomero 3,2020, Urupapuro 311-323.
Igihe cyo kohereza: Kanama-05-2021
