5' Cap保护mRNA不被外切酶降解，与3'端的poly A尾、poly A结合蛋白（PAPB）和翻译起始因子蛋白协同作用，起始蛋白质的翻译。体外转录（In Vitro Transcription，IVT）的加帽是常用的加帽方式，可以在IVT时加帽（即共转录加帽），也可以在IVT结束后加帽（即酶法加帽）【详见mRNA体外制备之酶法加帽和共转录加帽...
AChE mRNA STABILITY IN MAMMALIAN SKELETAL MUSCLE, STUDIED IN VITRO Institute of Pathophysiology, Medical Faculty, Ljubljana, SloveniaChim, ActaStabilitymammalian, R N AMuscle, SkeletalFaculty, Medical
2. Intradermal Delivery of Modified mRNA Encoding VEGF-A in Patients with Type 2 Diabetes; 3. mRNA-lipid nanoparticle COVID-19 vaccines: Structure and stability; 4. WO2012019780和US20150104476; 5. Protective efficacy of in vitro synthesized,specific messenger RNA vaccines against influenza A virus...
参考文献 [1] Kang DD, Li H, Dong Y. Advancements of in vitro transcribed mRNA (IVT mRNA) to enable translation into the clinics. Adv Drug Deliv Rev. 2023 Aug;199:114961. doi: 10.1016/j.addr.2023.114961. [2] Wayment-Steele HK, Kim DS, Choe CA, Das R, et al. Theoretical basis ...
5' Cap保护mRNA不被外切酶降解,与3'端的poly A尾、poly A结合蛋白(PAPB)和翻译起始因子蛋白协同作用,起始蛋白质的翻译。体外转录(In Vitro Transcription,IVT)的加帽是常用的加帽方式,可以在IVT时加帽(即共转录加帽),也可以在IVT结束后加帽(即酶法加帽)【详见mRNA体外制备之酶法加帽和共转录加帽】。
poly(A) tail对维持mRNA的稳定性起着重要作用,体外合成mRNA常用的加尾方式有两种:(1)在IVT(in vitro tranion)后利用poly (A) polymerase进行加尾;(2)质粒模板上设计poly (A)序列,在IVT时被转录出来从而带上poly (A)尾,由于成本低,工艺简便,这也是目前主流的方法。长片段的 poly(A)序列在质粒扩增过程中不稳...
此时,威廉·斯图蒂埃(William Studier)实验室克隆了T7 RNA聚合酶[25],并在1984年提交了专利,将其开发成一个高效且可诱导的体外转录系统(in vitrotranscription system)[26]。T7 RNA聚合酶具有几个有利特征,包括高度特异性地结合到T7启动子(相对于转录起始位点的保守核苷酸-17到+6位点)和高速转录RNA的能力。
Fig. 2:Excipients and buffer modifications for improved LNP stability and in vivo delivery. Fig. 3: Synthesis and screening of a biodegradable lipid library. Fig. 4: In vivo testing and in vitro–in vivo comparison of top lipids....
Fig. 1. Structural elements of in vitro transcribed (IVT) mRNA. Each of these elements can be optimized and modified in order to modulate the stability, translation capacity, and immune-stimulatory profile of mRNA. Courtesy of Verbeke et al. (2019). Many efforts have been made to increase ...
Challenges related to mRNA stability and immunogenicity, as well as in vivo delivery and the ability to cross multiple biological barriers, have been largely addressed by recent progress in mRNA engineering and delivery. In this Review, we present the latest advances and innovations in the growing ...