Overexpression of lncRNAs with endogenous lengths and functions using a lncRNA delivery system based on transposon.
Overexpression of lncRNAs with endogenous lengths and functions using a lncRNA delivery system based on transposon.
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使用基于转座子的lncRNA递送系统过度表达具有内源长度和功能的lncRNA
DOI：
10.1186/s12951-021-01044-7
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发表时间：
2021-10-02
影响因子：
10.2
通讯作者：
Yin D
中科院分区：
文献类型：
--
作者：
Zhang Y;Huang YX;Jin X;Chen J;Peng L;Wang DL;Li Y;Yao XY;Liao JY;He JH;Hu K;Lu D;Guo Y;Yin D
研究背景长链非编码RNA（lncRNA）在许多生理和病理过程中发挥重要作用，提示lncRNA可作为基因治疗的潜在靶点。稳定表达是lncRNA研究的基础技术。慢病毒是用于稳定表达的最广泛使用的递送系统之一。然而，它最初被设计为mRNA，和慢病毒载体的lncRNAs.ResultsWe发现，慢病毒载体产生lncRNAs与不正确的终止，附加一个额外的片段~ 2 kb的3 ′端的适用性是很大程度上unknown。结果表明，慢病毒载体并不是理想的lncRNA传递系统。在这里，我们开发了一种新的lncRNA递送方法，称为使用转座子系统表达具有内源性特征的lncRNA（ELECTS）。通过在lncRNA序列后插入终止信号，ELECTS产生没有3个侧翼序列的转录物，并保留lncRNA的天然特征和功能，这是慢病毒载体无法实现的。此外，ELECTS对操作员没有潜在的感染风险，并且所需时间更少。ELECTS为lncRNA.ConclusionsOur研究表明，慢病毒载体中不适当的转录终止对lncRNAs的分子作用和细胞功能有根本性的影响。本研究开发的ELECTS系统将为lncRNA的研究提供一种方便、可靠的方法。
BackgroundLong noncoding RNAs (lncRNAs) play important roles in many physiological and pathological processes, this indicates that lncRNAs can serve as potential targets for gene therapy. Stable expression is a fundamental technology in the study of lncRNAs. The lentivirus is one of the most widely used delivery systems for stable expression. However, it was initially designed for mRNAs, and the applicability of lentiviral vectors for lncRNAs is largely unknown.ResultsWe found that the lentiviral vector produces lncRNAs with improper termination, appending an extra fragment of ~ 2 kb to the 3ʹ-end. Consequently, the secondary structures were changed, the RNA–protein interactions were blocked, and the functions were impaired in certain lncRNAs, which indicated that lentiviral vectors are not ideal delivery systems of lncRNAs. Here, we developed a novel lncRNA delivery method called the Expression of LncRNAs with Endogenous Characteristics using the Transposon System (ELECTS). By inserting a termination signal after the lncRNA sequence, ELECTS produces transcripts without 3ʹ-flanking sequences and retains the native features and function of lncRNAs, which cannot be achieved by lentiviral vectors. Moreover, ELECTS presents no potential risk of infection for the operators and it takes much less time. ELECTS provides a reliable, convenient, safe, and efficient delivery method for stable expression of lncRNAs.ConclusionsOur study demonstrated that improper transcriptional termination from lentiviral vectors have fundamental effects on molecular action and cellular function of lncRNAs. The ELECTS system developed in this study will provide a convenient and reliable method for the lncRNA study.Graphic Abstract
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影响因子：
2.8
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
56.9
作者：
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通讯作者：
Wahlestedt, C