L17ER4: A cell-permeable attenuated cationic amphiphilic lytic peptide
L17ER4: A cell-permeable attenuated cationic amphiphilic lytic peptide
复制标题
L17ER4：细胞渗透性减毒阳离子两亲裂解肽
DOI：
10.1016/j.bmc.2022.116728
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Futaki Shiroh
中科院分区：
文献类型：
--
作者：
Shinga Kenta;Iwata Takahiro;Murata Kazuya;Daitoku Yoko;Michibata Junya;Arafiles Jan Vincent V.;Sakamoto Kentarou;Akishiba Misao;Takatani-Nakase Tomoka;Mizuno Seiya;Sugiyama Fumihiro;Imanishi Miki;Futaki Shiroh
我们已经开发了一系列减毒阳离子两亲性裂解（ACAL）肽，可以有效地将免疫球蛋白G（IgG）和其他功能蛋白带入细胞。递送通常通过ACAL肽与货物蛋白的共施用来实现。然而，ACAL肽与货物的缀合可能是一种有前途的体内应用方法，以连接ACAL肽和货物的体内结果。这项研究描述了一种新的细胞渗透性ACAL肽L17ER4的产生。L17 E是我们实验室先前开发的ACAL肽的优化原型，用于将IgG有效递送到细胞中。通过用四精氨酸（R4）标签官能化L17 E来改善递送。与使用R8（具有高细胞内递送功效的代表性细胞穿透肽）相比，与L17ER 4缀合提供模型小分子货物（异硫氰酸荧光素和HiBiT肽）的约4倍更高的细胞摄取。L17ER4还能够将蛋白质递送到细胞。将Cre重组酶与L17ER 4融合后导入细胞。将Cre-L 17 ER 4注射到绿色红色报告小鼠R26 GRR中，导致室管膜细胞中显著的体内基因重组，表明L 17 ER 4可用作细胞穿透肽，用于将蛋白质治疗剂递送到体内细胞中。
We have developed a series of attenuated cationic amphiphilic lytic (ACAL) peptides that can efficiently bring immunoglobulin G (IgG) and other functional proteins into cells. Delivery is generally achieved through the coadministration of ACAL peptides with cargo proteins. However, conjugation of ACAL peptides with cargos may be a promising approach forin vivoapplication to linkin vivooutcomes of ACAL peptides and cargos. This study describes the creation of a new cell-permeable ACAL peptide, L17ER4. L17E is an optimized prototype of ACAL peptides previously developed in our laboratory for efficient delivery of IgGs into cells. Delivery was improved by functionalizing L17E with a tetra-arginine (R4) tag. Compared to the use of R8, a representative cell-penetrating peptide with high intracellular delivery efficacy, conjugation with L17ER4 afforded approximately four-fold higher cellular uptake of model small-molecule cargos (fluorescein isothiocyanate and HiBiT peptide). L17ER4 was also able to deliver proteins to cells. Fused with L17ER4, Cre recombinase was delivered into cells. Intracerebroventricular injection of Cre-L17ER4 into green red reporter mice, R26GRR, led to significantin vivogene recombination in ependymal cells, suggesting that L17ER4 may be used as a cell-penetrating peptide for delivering protein therapeutics into cellsin vivo.