mRNA, lossis tus xa xov RNA, yog ib hom molecule uas nqa cov ntaub ntawv caj ces los ntawm DNA mus rau cov protein- ua tshuab ntawm tes. Los ntawm kev xa mRNA rau hauv cov hlwb, peb tuaj yeem qhia lawv kom tsim cov protein uas peb xav tau, xws li tshuaj tiv thaiv kab mob, enzymes, tshuaj hormones, lossis tshuaj tiv thaiv. Qhov no qhib lub ntiaj teb tshiab ntawm kev muaj peev xwm kho ntau yam kab mob, los ntawm cov kab mob sib kis mus rau mob qog noj ntshav.
Hauv tsab xov xwm no, peb yuav qhia koj txog cov hauv paus ntawm kev kho mRNA, suav nrog nws ua haujlwm li cas, nws ua li cas, nws xa li cas, thiab nws siv li cas. Peb kuj tseem yuav qhia qee qhov kev nce qib tshiab thiab cov kev sib tw hauv qhov chaw zoo siab ntawm biotechnology.
Kev kho mRNA ua haujlwm li cas?
mRNA kev kho mob ua haujlwm los ntawm kev qhia cov khoom siv mRNA molecules rau hauv cov hlwb, qhov chaw uas lawv tau muab txhais ua cov protein ua haujlwm. Nyob ntawm seb hom thiab lub hom phiaj ntawm cov protein, kev kho mRNA tuaj yeem muaj qhov sib txawv. Piv txwv li:
Cov tshuaj tiv thaiv mRNA tuaj yeem ua rau lub cev tsis muaj zog tiv thaiv kab mob tshwj xeeb, xws li tus kabmob tshiab tshiab uas ua rau COVID-19. Los ntawm kev xa cov mRNA uas encodes tus kab mob kis kab mob, xws li cov protein ntau, peb tuaj yeem txhawb kev tsim cov tshuaj tiv thaiv kab mob thiab T hlwb uas tuaj yeem paub thiab tshem tawm tus kab mob.
mRNA cov tshuaj tiv thaiv tuaj yeem muab kev tiv thaiv kab mob tiv thaiv kab mob, xws li rabies lossis HIV. Los ntawm kev xa cov mRNA uas encodes monoclonal antibody, peb tuaj yeem muab kev tiv thaiv tam sim ntawm ib qho antigen tshwj xeeb, yam tsis muaj kev cia siab rau lub cev lub cev tiv thaiv kab mob.
mRNA protein hloov tuaj yeem rov ua haujlwm ntawm cov protein uas ploj lawm lossis tsis zoo, xws li yam VIII hauv hemophilia lossis dystrophin hauv cov leeg nqaij dystrophy. Los ntawm kev xa mRNA uas encodes ib txwm version ntawm cov protein, peb tuaj yeem kho qhov ua rau tus kab mob thiab txhim kho cov tsos mob.
mRNA noob kho tuaj yeem hloov kho cov genome ntawm ib lub xovtooj ntawm tes, xws li kho qhov hloov pauv lossis ntxig cov noob tshiab. Los ntawm kev xa mRNA uas encodes programmable nucleases, xws li CRISPR/Cas9, peb tuaj yeem tsom thiab kho cov DNA ib ntus nrog kev ua haujlwm siab thiab ua haujlwm zoo.
mRNA ua li cas?
mRNA yog tsim los ntawm cov txheej txheem hu ua in vitro transcription (IVT), uas siv cov qauv DNA thiab RNA polymerase enzyme los ua ke RNA molecules. Cov qauv DNA muaj cov txheej txheem txhawb nqa thiab cov txheej txheem coding ntawm cov protein uas xav tau. RNA polymerase khi rau tus txhawb nqa thiab xa cov DNA rau hauv RNA.
Txhawm rau kom ntseeg tau tias cov hluavtaws mRNA ua haujlwm thiab ruaj khov hauv cov hlwb, ntau qhov kev hloov kho tau ua thaum lossis tom qab IVT. Cov no suav nrog:
Capping: ntxiv qhov hloov pauv nucleotide rau 5' kawg ntawm mRNA los tiv thaiv nws los ntawm kev degradation thiab txhim kho nws cov kev txhais lus zoo. Muaj ntau hom kaus mom, xws li Cap 0, Cap 1, thiab Cap 2, uas txawv ntawm lawv cov qib methylation.
Polyadenylation: ntxiv ib txoj hlua ntawm adenine nucleotides rau 3' kawg ntawm mRNA kom nws ruaj khov thiab ib nrab - lub neej. Qhov ntev ntawm poly(A) tus Tsov tus tw yuav txawv ntawm 10 mus rau 250 nucleotides.
Optimization: kho cov txheej txheem ntawm mRNA, xws li 5' untranslated region (UTR), 3' UTR, thiab qhib kev nyeem ntawv (ORF), los txhim kho nws cov theem qhia thiab kev ua. Qhov no yuav suav nrog kev ua kom zoo rau kev siv codon, qhia txog kev ua haujlwm peptides lossis cov teeb liab sib lawv liag, lossis ntxiv rau tus kheej - amplifying ntsiab.
Kev hloov kho: hloov kho cov qauv tshuaj ntawm qee cov nucleotides hauv mRNA kom txo nws txoj kev tiv thaiv kab mob thiab ua kom nws ruaj khov. Qhov no yuav suav nrog kev hloov uridine nrog pseudouridine lossis N1-methylpseuridine, lossis ntxiv methyl pawg rau lwm txoj haujlwm.
MRNA xa li cas?
mRNA kev xa khoom yog ib qho teeb meem loj tshaj plaws hauv kev kho mRNA, vim mRNA molecules loj, tsis zoo them, thiab nquag degradation los ntawm enzymes hauv cov kua roj ntsha. Yog li ntawd, ntau hom cab kuj tau tsim los tiv thaiv thiab thauj mRNA rau hauv cov hlwb. Cov no suav nrog:
Lipid nanoparticles (LNPs): spherical particles muaj lipids uas tsim ib tug bilayer nyob ib ncig ntawm mRNA molecules. LNPs tuaj yeem fuse nrog cell membranes thiab tso mRNA rau hauv cytoplasm. LNPs tau siv dav rau kev xa khoom ntawm mRNA vim lawv cov kev ua haujlwm siab thiab biocompatibility. Qee qhov piv txwv ntawm LNPs yog cationic solid lipid nanoparticles (SLNs), ionizable lipid nanoparticles (ILNs), thiab immunostimulatory lipid nanoparticles (ISLNs).
Polymer nanoparticles (PNPs): spherical particles muaj li ntawm polymers uas tsim ib tug tub ntxhais -plhaub qauv nyob ib ncig ntawm mRNA molecules. PNPs tuaj yeem nkag mus rau cov hlwb los ntawm endocytosis thiab khiav tawm ntawm lysosomes los ntawm cov nyhuv proton sponge lossis enzymatic degradation. PNPs muaj ntau yam thiab kho tau rau ntau daim ntawv thov. Qee qhov piv txwv ntawm PNPs yog polyethylene (PEI), poly (lactic-co-glycolic acid) (PLGA), thiab chitosan.
Cationic nanoemulsions (CNEs): tee roj-hauv- dej emulsions uas muaj cationic lipids thiab surfactants. CNEs tuaj yeem cuam tshuam nrog cell membranes thiab xa mRNA rau hauv cytoplasm. CNEs yooj yim los npaj thiab nce, thiab tuaj yeem txhim kho cov tshuaj tiv thaiv mRNA. Qee qhov piv txwv ntawm CNEs yog DOTAP / DOPE, DOTAP / cholesterol, thiab DOTAP / MCT.
Lwm lub tshuab xa khoom mRNA: qee qhov lwm hom kev thauj khoom uas tau tshawb xyuas rau mRNA xa khoom yog protamine-condensed mRNA, exosomes, extracellular vesicles (EVs), mesoporous silica, thiab calcium phosphate (CaP).
Dab tsi yog cov kev siv ntawm mRNA kev kho mob?
mRNA kev kho mob muaj peev xwm kho tau ntau yam kab mob, los ntawm cov kab mob sib kis mus rau cov kab mob metabolic, los ntawm mob qog noj ntshav mus rau cov kab mob plawv, thiab los ntawm cov kab mob caj ces mus rau cov tshuaj kho dua tshiab. Qee qhov kev siv tam sim no thiab tshwm sim ntawm kev kho mRNA yog:
Cov tshuaj tiv thaiv mRNA: cov tshuaj tiv thaiv mRNA tuaj yeem ua rau lub cev muaj zog thiab muaj zog tiv thaiv ntau yam kab mob, xws li SARS-CoV-2, kab mob khaub thuas, HIV, kab mob ua pa syncytial (RSV), herpes simplex virus (HSV), varicella zoster virus (VZV), human cyt virus (VZga), kab mob ntsws (VZga), kab mob ntsws (VZga) tus kab mob. Cov tshuaj tiv thaiv mRNA muaj ntau qhov zoo dua li cov tshuaj tiv thaiv ib txwm muaj, xws li kev txhim kho sai, tus nqi qis, kev nyab xeeb siab, thiab yooj yim scalability.
mRNA tshuaj tiv thaiv: mRNA cov tshuaj tiv thaiv tuaj yeem muab kev tiv thaiv kab mob tiv thaiv ntau yam kab mob, xws li rabies, HIV, Ebola, Zika, anthrax, thiab COVID-19. mRNA cov tshuaj tiv thaiv muaj ntau qhov zoo dua li cov tshuaj tiv thaiv kab mob, xws li kev qhia ntev, kev tiv thaiv kab mob qis, kev ruaj ntseg siab, thiab kev tswj xyuas yooj yim.
mRNA protein hloov: mRNA protein hloov tuaj yeem rov ua haujlwm ntawm cov protein uas ploj lawm lossis tsis zoo hauv ntau yam kab mob, xws li hemophilia, muscular dystrophy, cystic fibrosis, phenylketonuria, Fabry disease, Gaucher disease, thiab Parkinson's disease. mRNA hloov pauv protein muaj ntau qhov zoo ntawm kev kho cov protein, xws li zam kev tiv thaiv kab mob, txhim kho bioavailability, thiab txo qhov ntau npaum li cas thiab ntau zaus.
mRNA gene editing: mRNA gene editing tuaj yeem hloov kho cov genome ntawm cov cell hauv ntau yam kab mob, xws li mob cell anemia, beta-thalassemia, Huntington's disease, Duchenne muscular dystrophy, thiab cystic fibrosis. mRNA gene editing muaj ntau yam zoo dua DNA-raws li kev kho noob caj noob ces, xws li zam kev koom ua ke rau hauv tus tswv tsev genome, txo tawm - cov phiaj xwm cuam tshuam, thiab nce qhov tshwj xeeb thiab kev ua tau zoo.
mRNA cell therapy: mRNA cell therapy tuaj yeem tswj cov haujlwm ntawm lub xovtooj ntawm ntau yam kab mob, xws li mob qog noj ntshav, kab mob autoimmune, ntshav qab zib, thiab kab mob plawv. mRNA cell therapy muaj ntau yam zoo dua li cov kev kho ntawm tes, xws li tsis txhob kis kab mob, txhim kho kev nyab xeeb thiab ntau yam, thiab pab txhawb kev tsim khoom thiab kev tswj kom zoo.
Dab tsi yog cov teeb meem thiab cov cib fim ntawm kev kho mRNA?
mRNA kev kho mob yog ib qho kev cog lus thiab tawm ntawm biotechnology uas tau pom muaj peev xwm kho tau ntau yam kab mob. Txawm li cas los xij, tseem muaj qee qhov kev sib tw thiab kev txwv uas yuav tsum tau kov yeej ua ntej nws tuaj yeem siv dav hauv kev kho mob. Cov no suav nrog:
- Txhim kho qhov ruaj khov thiab kev xa tawm ntawm mRNA molecules hauv vivo
-Txo cov immunogenicity thiab toxicity ntawm mRNA molecules thiab cov cab kuj
-Kev ua kom zoo ntawm qib kev qhia thiab lub sijhawm ntawm mRNA-encoded proteins
-Tsim cov txheej txheem kev cai thiab kev tswj xyuas zoo rau kev tsim mRNA
- Ntsuas qhov ntev - kev nyab xeeb thiab kev ua tau zoo ntawm kev kho mRNA hauv tib neeg
-Txawm hais tias cov kev cov nyom no, kuj tseem muaj ntau lub cib fim thiab qhov zoo uas ua rau kev kho mRNA yog qhov kev xaiv zoo rau cov tshuaj yav tom ntej. Cov no suav nrog:
-Kev siv qhov hloov tau yooj yim thiab ntau haiv neeg ntawm mRNA tsim thiab kev sib txuas
-Leveraging inherent immunogenicity thiab adjuvanticity ntawm mRNA molecules
-Kev siv cov kev qhia tsis tu ncua thiab rov qab los ntawm mRNA-encoded proteins
- Tshawb nrhiav cov txiaj ntsig kev sib koom ua ke ntawm kev sib koom ua ke mRNA kev kho mob nrog lwm yam kev hloov kho
-Kev nrawm nrawm ntawm kev txhim kho thiab kev pom zoo ntawm mRNA cov khoom
Prisys Biotechnology, lub tuam txhab tshwj xeeb hauvtsis yog-kev tshawb fawb tib neeg primate (NHP).thiab cov kev pabcuam rau kev txhim kho tshuaj preclinical. mRNA therapy, ib hom molecule uas tuaj yeem qhia cov hlwb los tsim cov protein uas peb xav tau, xws li tshuaj tiv thaiv kab mob, enzymes, tshuaj hormones, lossis tshuaj tiv thaiv. Cynomolgus macaques, ib hom ntawm NHP uas muaj ntau yam zoo sib xws nrog tib neeg nyob rau hauv cov ntsiab lus ntawm lub cev, lub cev nqaij daim tawv, kev tiv thaiv kab mob, thiab noob caj noob ces. Prisys Biotechnology hauv mRNA tshuaj tshawb fawb thiab kev loj hlob thiab NHP qauv raws li hauv qab no:
Prisys Biotechnology muaj kev sib koom ua ke ntawm NHP cov peev txheej, cov qauv kab mob, thiab cov tsev kho mob kho mob los muab kev daws teeb meem rau ntau yam kab mob, xws li kev tiv thaiv kab mob, mob, kab mob plawv, hematology, siab, raum, ntsws, thiab oncology.
Prisys Biotechnology muaj kev koom tes tsim kev xaiv rau R&D kev sib koom tes thiab cov thev naus laus zis tshiab xws li kev kawm tob -raws li 3D kev coj tus cwj pwm kom muaj nuj nqis hauv NHP.
Prisys Biotechnology siv cynomolgus macaques ua tus qauv tsiaj rau kev kho mRNA, uas muaj kev ntseeg siab thiab cuam tshuam ntau dua li lwm cov qauv tsiaj, xws li nas lossis nas.
Prisys Biotechnology tuaj yeem kuaj qhov muaj peev xwm thiab muaj peev xwm ntawm kev kho mRNA rau ntau yam kab mob los ntawm kev ntsuas cov hluav taws xob tiv thaiv, cov khoom sib nqus, cov tshuaj pharmacokinetics, toxicity, immunogenicity, thiab kev ua tau zoo ntawm mRNA molecules thiab cov neeg nqa khoom hauv NHPs.
