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肽偶联 27 个核苷酸小干扰 RNA 增强基因沉默效果和膜通透性。

Enhancement of gene silencing effect and membrane permeability by Peptide-conjugated 27-nucleotide small interfering RNA.

机构信息

Faculty of Pharmacy, Yasuda Women's University, 6-13-1 Yasuhigashi, Asaminami-ku, Hiroshima 731-0153, Japan.

出版信息

Molecules. 2012 Sep 14;17(9):11089-102. doi: 10.3390/molecules170911089.

DOI:10.3390/molecules170911089
PMID:22983148
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6268710/
Abstract

Two different sizes of siRNAs, of which one type was 21-nucleotide (nt) siRNA containing 2-nt dangling ends and the other type was 27-nt siRNA with blunt ends, were conjugated with a nuclear export signal peptide of HIV-1 Rev at the 5'-sense end. Processing by Dicer enzyme, cell membrane permeability, and RNAi efficiency of the peptide-conjugated siRNAs were examined. Dicer cleaved the peptide-conjugated 27-nt siRNA leading to the release of 21-nt siRNA, whereas the peptide-conjugated 21-nt siRNA was not cleaved. High membrane permeability and cytoplasmic localization was found in the conjugates. Moreover, the peptide-conjugated 27-nt siRNA showed increased potency of RNAi in comparison with the nonmodified 21-nt and 27-nt siRNAs, whereas the peptide-conjugated 21-nt siRNA showed decreased RNAi efficacy. This potent RNAi efficacy is probably owing to acceleration of RISC through recognition by Dicer, as well as to the improvement of cell membrane permeability and intracellular accumulation.

摘要

两种不同大小的 siRNA，其中一种是 21 个核苷酸（nt）的 siRNA，含有 2 个 nt 的悬垂末端，另一种是 27 个核苷酸的 siRNA，具有钝末端，它们都在 5'-感测末端与 HIV-1 Rev 的核输出信号肽连接。检查了肽偶联 siRNA 的 Dicer 酶处理、细胞膜通透性和 RNAi 效率。Dicer 酶切割肽偶联的 27-nt siRNA，导致释放 21-nt siRNA，而肽偶联的 21-nt siRNA未被切割。在缀合物中发现了高的膜通透性和细胞质定位。此外，与未修饰的 21-nt 和 27-nt siRNA 相比，肽偶联的 27-nt siRNA 显示出增强的 RNAi 效力，而肽偶联的 21-nt siRNA 显示出降低的 RNAi 效力。这种有效的 RNAi 效力可能归因于通过 Dicer 的识别加速了 RISC，以及改善了细胞膜通透性和细胞内积累。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/007b/6268710/39a3f651724c/molecules-17-11089-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/007b/6268710/79520625346c/molecules-17-11089-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/007b/6268710/37ffe0fff778/molecules-17-11089-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/007b/6268710/d4b5e938ab5a/molecules-17-11089-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/007b/6268710/62bc1e5281db/molecules-17-11089-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/007b/6268710/39a3f651724c/molecules-17-11089-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/007b/6268710/79520625346c/molecules-17-11089-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/007b/6268710/37ffe0fff778/molecules-17-11089-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/007b/6268710/d4b5e938ab5a/molecules-17-11089-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/007b/6268710/62bc1e5281db/molecules-17-11089-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/007b/6268710/39a3f651724c/molecules-17-11089-g004.jpg

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