2018年诺华公司Beverly M等发表在Anal Bioanal Chem杂志的一篇文章，《Poly A tail length analysis of in vitro transcribed mRNA by LC-MS》，研究基于RNAse T1酶切与oligo dT磁珠纯化得到polyA尾，使用液相色谱质谱联用（LC-MS）检测polyA尾长度的分布，目前该策略是当前mRNA体外制备工艺中较为常用的方法。2022年...
基于核酸酶切割的poly A尾长度分析策略使用较为广泛,使用RNase T1将mRNA 3’端polyA切割纯化,然后通过高分辨LC-MS分离鉴别不同长度的polyA序列。 2.1酶切处理 据2018年诺华公司Beverly M等发表的文章:《Poly A tail length analysis of in vitro transcribed mRNA by LC-MS》,研究人员基于RNAse T1酶切与oligo ...
但当poly(A)尾长度不足16nt时,mRNA无法翻译,而且胞内很少观察到poly(A)尾长度在30nt以下的mRNA【45】。上述证据综合起来说明poly(A)尾稳定了mRNA(允许其翻译),30nt通常是mRNA实现稳定性的下限,而这正好是PABPC对poly(A)尾长度的最低要求。但凡事都有例外,带有极短(小于20nt)poly(A)尾甚至不含poly(A)尾...
三、Poly(A)尾长度、翻译效率和mRNA稳定性之间并不是简单的线性关系。高度翻译的mRNA具有约30个腺苷的短poly(A)尾，该长度只能容纳单个PABPC，这表明一个PABPC足以促进有效翻译。Poly(A)尾作为mRNA翻译和稳定性的中心调节剂，将随着mRNA的治疗手段和疫苗的兴起成为该领域的研究热潮。因此理解poly(A)尾在基因表达调...
3'-End processing of pre-mRNA in eukaryotes. NA Degradation in Saccharomyces cerevisae. The Human Nuclear Poly(A)-Binding Protein Promotes RNA Hyperadenylation and Decay. Role of poly (A) tail as an identity element for mRNA nuclear export. ...
据2018年诺华公司Beverly M等发表的文章:《Poly A tail length analysis of in vitro transcribed mRNA by LC-MS》,研究人员基于RNAse T1酶切与oligo dT磁珠纯化得到3’端polyA尾,使用液相色谱质谱联用(LC-MS)检测polyA尾长度的分布。 具体步骤包括:RNase T1酶切、mRNA 3’端polyA磁珠纯化(oligo dT)。RNase T1...
[8] Lai WC, Zhu M, Belinite M, Ermolenko DN, et al. Intrinsically Unstructured Sequences in the mRNA 3' UTR Reduce the Ability of Poly(A) Tail to Enhance Translation. J Mol Biol. 2022 Dec 30;434(24):167877. doi: 10.1016/j.jmb.2022.167877. ...
mRNA, PolyA Tail, OocytesLin, R
Beverly M等人(“Poly A tail length analysis of in vitro transcribedmRNA by LC-MS”,《Anal Bioanal Chem》,2018年)提出了一种定量分析mRNA的polyA尾长度的方法,该方法基于内切酶RNase T1可专一性地在单链RNA的鸟嘌呤核糖核苷酸(G)以及3’端相邻核糖核苷酸间进行切割磷酸二酯键,通过polyA尾与oligo dT磁珠特异...
mRNA aftertranscription, helped export the mRNA to the cytoplasm (Darnellet al.1971). Once the mRNA got into the cytoplasmic space, the shorter poly(A) tail was not sufficient to permitproteintranslation. It turns out that the length of the poly(A) tails in the cytoplasm is extremely ...