DNA ndi RNA kaphatikizidwe zimachokera pa njira yolimba-gawo kaphatikizidwe ndi phosphoramidite chemistry, DNA synthesizer angagwiritsidwe ntchito synthesize RNA kapena RNA analogue popanda kusinthidwa kwina, ndi reagents mu DNA kaphatikizidwe angagwiritsidwe ntchito mwachindunji mu RNA ndi yokumba nucleic acid. ' synthesis.Monga momwe tawonetsera pa Chithunzi 1, 2'-hydroxy mu RNA phosphoramidite imatetezedwa ndi gulu loteteza la silyl, mwachitsanzo t-butyldimethylsilyl (TBDMS), kuteteza zotsatira za mbali pa gulu losalimba la 2'-hydroxy.Gulu lalikulu la TBDMS lidalepheretsa zomwe zimachitika pakati pa phosphoramidite ndi gulu la 5'-hydroxy pa chithandizo cholimba, ndipo nthawi yayitali yolumikizira imafunikira kuti mukwaniritse kugwirizanitsa bwino.
Zofanana ndi kaphatikizidwe ka DNA, RNA yopangidwa idadulidwa koyamba kuchokera ku chithandizo cholimba ndi aminolysis, kenako gulu la TBDMS lidadulidwa ndi tetrabutylammonium fluoride (TBAF) kapena trimethylamine trihydrofluoride.Crude RNA ikhoza kuyeretsedwa ndi kukonzanso kuchokera ku mowa ndi HPLC.
Chithunzi 1. Kapangidwe kakemidwe kazinthu zomangira mu DNA ndi kaphatikizidwe ka RNA.
a) dABz phosphoramidite ndi b) rABz 2'-OTBDMS phosphoramidite.
Kukula kwa mankhwala a siRNA kumafuna kuti ma analogue amtundu wa RNA awonjezere kuyanjana, 2'-hydroxy pa RNA ikhoza kusinthidwa ndi gulu la MeO, F ndi MOE, ndipo nucleic acid yotsekedwa (LNA) imaperekanso ntchito yabwino mu RNA therapy. (Chithunzi 2).Ma phosphoramidite awa amapereka ntchito zofanana ndi DNA mtundu wa phosphoramidites mu kaphatikizidwe, ndi ntchito-mmwamba ndi kuyeretsa ma nucleic acid amenewa sanali achilengedwe ndi ofanana ndi DNA mbadwa.
Chithunzi 2. Kapangidwe kake kazinthu zomangira mu siRNA mankhwala.a) dABz 2-MeO phosphoramidite;b) dABz 2-F phosphoramidite;c) dABz 2-MOE phosphoramidite ndi d) dABz Yotsekedwa phosphoramidite.
Phosphoramiditesndi zofunika kuti kaphatikizidwe majini, kuphatikizapo makamaka DNA ndi RNA mabanja ndi zotumphukira awo, phosphoramidite onse pamwamba tikhoza kupereka mu phukusi.
Tikuwongolera magwiridwe antchito azinthu zathu nthawi zonse, kukonza njira zathu zopangira komanso kupeza tsatanetsatane wolondola.Sitimakupatsirani zinthu zapamwamba zokha, komanso timakupatsirani maphunziro ndi ntchito.Tili ndi antchito oyang'anira akatswiri komanso gulu labwino kwambiri laukadaulo ndi ogwira ntchito yokonza kuti tipatse makasitomala athu ntchito yabwino ikatha kugulitsa.
Nthawi yotumiza: Aug-09-2022