Kodi katemera wa mRNA ndi chiyani
Katemera wa mRNA amasamutsa RNA kupita ku maselo amthupi kuti afotokoze ndikupanga ma antigen a protein pambuyo pakusintha koyenera mu vitro, motero amatsogolera thupi kuti lipange chitetezo chamthupi motsutsana ndi antigen, potero kumakulitsa chitetezo chamthupi.[1,3].
Chithunzi 1: Chithunzi chojambula cha zotsatira za jakisoni wachindunji wa katemera wa mRNA [2]
Gulu la katemera wa mRNA
Katemera wa mRNA agawidwa m'mitundu iwiri:zosabwerezabwerezamRNA ndikudzikuzamRNA: kudzikuza mRNA sikungoyika ma antigen omwe mukufuna, komanso kumangiriza kubwereza komwe kumathandizira kukulitsa kwa RNA ndi njira yowonetsera mapuloteni.Makatemera osabwerezabwereza a mRNA amangoyika ma antigen omwe akuwatsata ndipo amakhala ndi zigawo 5'ndi 3' zosatembenuzidwa (UTR).Amapereka kukondoweza kwathunthu kwa kusinthika ndi chitetezo chamthupi, chomwe ndi mawu a antigen mu situ ndi kufalitsa chizindikiro chowopsa, ndipo ali ndi izi[2,3]
● Itha kupereka chilimbikitso chokwanira cha kusinthika ndi chitetezo chamthupi, chomwe ndi mawu a antigen mu situ ndi kufalitsa chizindikiro chowopsa.
●Zitha kupangitsa kuti chitetezo cha mthupi chitetezeke "choyenera", kuphatikizapo zochititsa chidwi komanso zamtundu wa cellular komanso chitetezo chamthupi
● Atha kuphatikiza ma antigen osiyanasiyana popanda kuwonjezera zovuta za kupanga katemera
● Kupititsa patsogolo mphamvu za chitetezo cha mthupi kumatha kutheka kudzera mu katemera mobwerezabwereza, ndipo palibe kapena kuchepa kwa chitetezo cha mthupi kwa wonyamulirayo.
●Katemera wosasunthika wa mRNA amathandizira kunyamula ndi kusunga katemera mosavuta
Chithunzi 2: Chithunzi chojambula cha katemera wa mRNA ndi kachitidwe kake ka antigen [4]
Zotsatira za katemera wa mRNA
Poyerekeza ndi katemera wakale, katemera wa mRNA ali ndi njira zosavuta zopangira, kufulumira kwachitukuko, kusowa kwa chikhalidwe cha maselo, komanso mtengo wotsika.Poyerekeza ndi katemera wa DNA, katemera wa mRNA safunikira kulowa mkati ndipo palibe chiwopsezo chophatikizana ndi ma genome omwe ali nawo.Theka la moyo likhoza kusinthidwa ndi kusinthidwa.
Gulu 1: Ubwino ndi kuipa kwa katemera wa mRNA
|
| Ubwino | Kuperewera |
| katemera wa mRNA | Kafukufuku wofulumira ndi chitukuko, kupanga katemera kumatenga masiku 40 okha | Yambitsani kuyankha kosayenera kwa chitetezo chamthupi
|
| Kusakhazikika kwa mRNA pansi pazikhalidwe zakuthupi, zosavuta kuzichepetsa | Sizingaphatikizidwe mu ma genome kuti mupewe kusintha kwachirengedwe kotheka
| |
| Palibe chifukwa cha chizindikiro chilichonse cha nyukiliya, zolembedwa | Mphamvu ya chitetezo cha nyukiliya ikuyenera kutsimikiziridwa
|
Chithunzi 3: Tchati choyenda cha katemera wa mRNA kupanga ndi kukonzekera [4]
Foregene Viral RNA Isolation kit
RT-qPCR Easy (Chigawo Chimodzi)
Njira zowongolera zopangira katemera wa mRNA
Chifukwa cha kusakhazikika bwino kwa mRNA yokha, kuwonongeka kosavuta ndi ma nucleases mu minofu, kutsika kwa maselo a cell, komanso kumasulira kosavuta, zolakwikazi zimachepetsa kugwiritsa ntchito katemera wa mRNA.Kumasulira bwino kumagwiranso ntchito yofunika kwambiri.Magalimoto operekera amatha kugawidwa kukhala ma vector a ma virus ndi ma vector omwe alibe ma virus (kuphatikiza ma liposomes, non-liposomes, virus, nanoparticles, etc.).Chifukwa chake, njira zoyenera zowongolera ndizofunikira.Zotsatirazi ndi njira yosinthira mankhwala pokonzekera mRNA[2]
1 Phatikizani ma analogi a kapu kapena gwiritsani ntchito ma enzymes kuti mukhazikitse mRNA ndikuwonjezera kumasulira kwa mapuloteni pomangirira ku eukaryotic translation initiation factor 4E (EIF4E)
2 Sinthani zinthu m'chigawo cha 5'-untranslated (UTR) ndi 3'-UTR kuti mukhazikike mRNA ndikuwonjezera kumasulira kwa mapuloteni
3 Kuwonjezera Poly(A) mchira kumatha kukhazikika mRNA ndikuwonjezera kumasulira kwa mapuloteni
4 Ma nucleosides osinthidwa kuti achepetse kuyambika kwa chitetezo chamthupi ndikuwonjezera kumasulira
5 Kuchiza ndi RNase III ndi kuyeretsa kwa protein yamadzimadzi chromatography (FPLC) kumatha kuchepetsa kutsegulira kwa chitetezo chamthupi ndikuwonjezera kumasulira.
6 Konzani masanjidwe kapena ma codon kuti muwonjezere zomasulira
7 Kupereka limodzi zinthu zoyambitsira zomasulira ndi njira zina zosinthira kumasulira ndi kutetezedwa
Chithunzi 4: In vitro transcription (IVT) mRNA kupanga ndi kusonkhana [5]
Kukonzekera kwakukulu kwa plasmid DNA
Kuyeretsedwa kwa plasmid DNA kumachotsa makamaka zowononga monga RNA, DNA endotoxin yotseguka, mapuloteni okhala ndi nucleic acid, ndipo nthawi zambiri amasintha plasmid yophatikizana kukhala E. coli.E. coli imalowa mkati mwa fermentation yochuluka kwambiri, kenako imasiyanitsidwa ndi madzi olimba, ndi kusonkhanitsa E. coli.E. coli ndiye pansi pa alkaline lysis, centrifugal olimba-zamadzimadzi kulekana ndi microfiltration kumveka pambuyo lysis, ultrafiltration ndi ndende pambuyo kufotokoza, ndiyeno chromatographic kuyeretsedwa.
Kuyeretsedwa kwa plasmid DNA:
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.Katemera wa mRNA - nyengo yatsopano mu katemera[J].Ndemanga Zachilengedwe Kupezeka Kwamankhwala, 2018.
【3】Kramps T., Elbers K. (2017) Mau oyamba a RNA Vaccines.Mu: Kramps T., Elbers K. (eds) RNA Vaccines.Njira mu Biology ya Molecular, vol 1499. Humana Press, New York, NY.
【4】Maruggi G, Zhang C, Li J, et al.mRNA monga Transformative Technology for Vaccine Development to Control Infectious Diseases[J].Chithandizo cha Molecular, 2019.
【5】Sergio Linares-Fernández, Céline Lacroix, ,Tailoring mRNA Vaccine to Balance Innate/Adaptive Immune Response,Trends in Molecular Medicine,Volume 26, Issue 3,2020,Masamba 311-323.
Nthawi yotumiza: Aug-05-2021
