Gịnị bụ ọgwụ mgbochi mRNA
Ogwu mRNA na-ebufe RNA n'ime sel nke ahụ iji gosipụta ma mepụta antigens protein mgbe mgbanwe dị mkpa na vitro, si otú a na-eduga anụ ahụ ka ọ mepụta nzaghachi mgbochi megide antigen, si otú ahụ gbasaa ikike nke ahụ.[1,3].
Onyonyo 1: Eserese atụmatụ nke mmetụta nke ịgba ọgwụ mgbochi mRNA kpọmkwem [2]
Nhazi nke ọgwụ mgbochi mRNA
A na-ekewa ọgwụ mgbochi mRNA ụzọ abụọ:enweghị atụmRNA nana-amụba onwe yamRNA: mRNA na-ebuli onwe ya ọ bụghị nanị na-edobe antigen e lekwasịrị anya, kamakwa ọ na-edokwa ngbanwe nke na-eme ka mmụba RNA intracellular na usoro okwu protein.Ọgwụ mgbochi mRNA na-anaghị emegharị ahụ na-edobe naanị antigens ebumnuche ma nwee mpaghara 5'na 3' asụgharịghị ya (UTR).Ha na-enye mkpali zuru oke nke ngbanwe na mgbochi ebumpụta ụwa, ya bụ na okwu antigen situ na nnyefe ihe egwu, ma nwee ngwa ndị a Atụmatụ.[2,3]
●nwere ike inye mkpali zuru oke nke mgbanwe na mgbochi ebumpụta ụwa, ya bụ na nkwupụta antigen situ na nnyefe mgbaàmà ihe egwu.
●nwere ike ịkpata mmeghachi omume mgbochi "kwesịrị ekwesị", gụnyere ihe ọchị na mmetụta cellular yana ebe nchekwa nchekwa
● Nwere ike ijikọ antigens dị iche iche na-enweghị ịbawanye mgbagwoju anya nke imepụta ọgwụ mgbochi
● Enwere ike imeziwanye ikike nke usoro ahụ ji alụso ọrịa ọgụ site n'ịgba ọgwụ mgbochi ugboro ugboro, ọ nweghịkwa ma ọ bụ obere nzaghachi mgbochi na-enye ndị na-ebu ya.
●Ọgwụ mgbochi mRNA na-ekpo ọkụ nwere ike ime ka njem na nchekwa nke ọgwụ mgbochi dị mfe
Onyonyo 2: Eserese atụmatụ nke ọgwụ mgbochi mRNA na usoro okwu antigen ya [4]
Atụmatụ ọgwụ mgbochi mRNA
Tụnyere ọgwụ mgbochi ọdịnala, ọgwụ mgbochi mRNA nwere usoro mmepụta dị mfe, ngwa ngwa mmepe ngwa ngwa, enweghị mkpa maka ọdịbendị cell, yana ọnụ ala dị ala.Tụnyere ọgwụ mgbochi DNA, ọgwụ mgbochi mRNA adịghị mkpa ịbanye n'ime oghere ma ọ nweghị ihe ize ndụ nke itinye n'ime genome nnabata.Enwere ike imezi ọkara ndụ site na mgbanwe.
Tebụl 1: Uru na ọghọm dị na ọgwụ mgbochi mRNA
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| Uru | adịghị mkpirisi |
| ọgwụ mgbochi mRNA | Nnyocha na mmepe ngwa ngwa, mmepụta ọgwụ mgbochi na-ewe naanị ụbọchị 40 | kpalite nzaghachi mgbochi ọrịa na-enweghị isi
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| Akwụsịghị mRNA n'okpuru ọnọdụ physiological, dị mfe imebi | Agaghị abanye na genome iji zere mmụgharị ọgwụgwọ enwere ike
| |
| Enweghị mkpa maka mgbaama mpaghara mpaghara nuklia ọ bụla, ndegharị | A ka ga-enyocha ịdị irè nke nuklia nchekwa
|
Foto 3: eserese ngosi nke mmepụta na nkwadebe ọgwụ mgbochi mRNA [4]
Ngwa Mwepu RNA nke Foregene
RT-qPCR dị mfe (Otu nzọụkwụ)
Atụmatụ emelitere maka ịkwado ọgwụ mgbochi mRNA
N'ihi nkwụsi ike na-adịghị mma nke mRNA n'onwe ya, nbibi dị mfe site na nucleases na anụ ahụ, arụmọrụ ntinye cell dị ala, na arụmọrụ ntụgharị dị ala, ntụpọ ndị a na-egbochi itinye ọgwụ mgbochi mRNA.Ịrụ ọrụ ntụgharị asụsụ na-arụkwa ọrụ dị oke egwu.Enwere ike kewaa ụgbọ ala ebufe ya na vectors na-abụghị nke nje (gụnyere liposomes, non-liposomes, virus, nanoparticles, wdg).Ya mere, a chọrọ usoro nkwalite ndị dị mkpa.Ihe a bụ atụmatụ nkwalite ọgwụ maka nkwadebe mRNA[2]
1 Synthesize okpu analọg ma ọ bụ jiri capping enzymes kwụsie ike mRNA ma welie ntụgharị asụsụ protein site na ijikọ ihe mmalite mmalite ntụgharị eukaryotic 4E (EIF4E)
2 Hazie ihe ndị dị na mpaghara 5′-enweghị ntụgharị (UTR) na 3′-UTR iji mee ka mRNA kwụsie ike ma welie ntụgharị asụsụ protein.
3 Ịgbakwunye Poly(A) ọdụ nwere ike kwụsie ike mRNA ma welie ntụgharị asụsụ protein
4 Nucleosides megharịrị ka o wedata n'ọgbalite mgbochi ọrịa ma bulie ntụgharị asụsụ
5 Ọgwụgwọ na RNase III na protein ngwa ngwa chromatography (FPLC) dị ọcha nwere ike ibelata ịgbalite mgbochi ọrịa yana ịbawanye ntụgharị asụsụ.
6 Kwalite usoro ma ọ bụ codons ka iwelie ntụgharị asụsụ
7 Ntinye aka nke ihe mmalite ntụgharị asụsụ na ụzọ ndị ọzọ iji gbanwee ntụgharị asụsụ na immunogenicity
Ọgụgụ 4: In vitro transcription (IVT) mRNA mmepụta na mgbakọ usoro [5]
Nkwadebe buru ibu nke DNA plasmid
Plasmid DNA dị ọcha na-ebutekarị mmetọ dị ka RNA, endotoxin DNA mepere emepe, protein host na host nucleic acid, ma na-agbanwekarị plasmid recombinant ka ọ bụrụ E. coli.E. coli na-enweta gbaa ụka njupụta dị elu, mgbe ahụ nkewa nke mmiri siri ike, na nchịkọta E. coli.A na-edobe E. coli na alkaline lysis, centrifugal siri ike-mmiri nkewa na microfiltration nkọwa mgbe lysis, ultrafiltration na itinye uche mgbe nkọwa, na mgbe ahụ chromatographic ọcha.
Nhicha nke DNA plasmid:
Foregene General 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 , et al.ọgwụ mgbochi mRNA - oge ọhụrụ na ịgba ọgwụ mgbochi ọrịa [J].Nleba anya nke okike nchọpụta ọgwụ, 2018.
【3】Kramps T., Elbers K. (2017) Okwu Mmalite nke RNA Vaccines.Na: Kramps T., Elbers K. (eds) Ọgwụ mgbochi RNA.Ụzọ na Molecular Biology, vol 1499. Humana Press, New York, NY.
【4】Maruggi G , Zhang C , Li J , et al.mRNA dị ka teknụzụ mgbanwe maka mmepe ịgba ọgwụ mgbochi iji chịkwaa ọrịa na-efe efe[J].Therapy Molecular, 2019.
【5】Sergio Linares-Fernández, Céline Lacroix, ,Tailoring mRNA Vaccine to Balance Innate/Adaptive Immune Response,Trends in Molecular Medicine,Volu 26, Issue 3,2020,page 311-323.
Oge nzipu: Ọgọst-05-2021
