Mechanisms and Strategies for Enhancing DNA Nuclear Entry in Gene Delivery.

IF 3.5 4区医学 Q3 BIOPHYSICS

Cellular and molecular bioengineering Pub Date : 2026-06-29 DOI:10.1007/s12195-026-00927-9

Justin Sylvers, Fan Yuan

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引用次数: 0

Abstract

Background: Nonviral gene delivery using DNA vectors is widely used in cell engineering, vaccination, and gene therapy, but delivery efficiency remains lower than those of viral vectors and mRNA-based approaches, partly due to inefficient nuclear entry, as transfected DNA must enter the nucleus for transcription. Therefore, a mechanistic understanding of nuclear entry pathways is essential for developing strategies to improve the efficiency.

Methods: This review evaluated mechanistic studies of DNA nuclear entry in mammalian cells, mathematical models of intracellular DNA trafficking, quantitative analyses of DNA nuclear accumulation and transgene expression, and strategies to enhance nuclear delivery of DNA.

Results: Two mechanistically distinct pathways for DNA nuclear entry have been reported: enclosure upon nuclear envelope reformation in dividing cells, and active transport through nuclear pore complexes (NPCs). Various strategies have been developed to enhance nuclear import through these pathways; however, their effectiveness depends on multiple factors, including cell type, delivery methods, and cell cycle status. Although DNA vectors are significantly larger than the nominal inner diameter of NPCs, they may traverse NPCs through deformation and interactions with nuclear transport proteins. Quantitative studies show that DNA nuclear accumulation is time dependent and heterogeneous among individual cells within the same population.

Conclusions: Nuclear entry plays a key role in determining efficiency of nonviral gene delivery. Advances in mechanistic studies, quantitative modeling, and imaging-based analyses have improved our understanding of intracellular DNA trafficking and nuclear accumulation. Integrating these insights with delivery strategies that enhance nuclear access while preserving the cellular machinery required for transgene expression will be critical for developing more efficient and reliable nonviral DNA delivery systems for therapeutic and biotechnological applications.

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基因传递中增强DNA核进入的机制和策略。

背景：使用DNA载体的非病毒基因传递广泛应用于细胞工程、疫苗接种和基因治疗，但传递效率仍然低于病毒载体和基于mrna的方法，部分原因是由于转染的DNA必须进入细胞核进行转录，因此传递效率较低。因此，对核进入途径的机制理解对于制定提高效率的策略至关重要。方法：本文综述了哺乳动物细胞内DNA进入细胞核的机制、细胞内DNA运输的数学模型、DNA核积累和转基因表达的定量分析以及增强DNA核递送的策略。结果：DNA进入细胞核有两种不同的机制途径：分裂细胞中核膜重组时的封闭，以及通过核孔复合物（NPCs）的主动运输。已经制定了各种战略，通过这些途径加强核进口；然而，它们的有效性取决于多种因素，包括细胞类型、给药方式和细胞周期状态。尽管DNA载体明显大于npc的名义内径，但它们可能通过变形和与核转运蛋白的相互作用穿越npc。定量研究表明，DNA核积累是时间依赖性的，并且在同一种群内的单个细胞之间具有异质性。结论：核进入在非病毒基因传递效率中起关键作用。机制研究、定量建模和基于成像的分析的进展提高了我们对细胞内DNA运输和核积累的理解。将这些见解与递送策略结合起来，既能增强核通路，又能保留转基因表达所需的细胞机制，对于开发更有效、更可靠的非病毒DNA递送系统，用于治疗和生物技术应用至关重要。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Cellular and molecular bioengineering 生物-生物物理

CiteScore

5.60

自引率

3.60%

发文量

审稿时长

>12 weeks

期刊介绍： The field of cellular and molecular bioengineering seeks to understand, so that we may ultimately control, the mechanical, chemical, and electrical processes of the cell. A key challenge in improving human health is to understand how cellular behavior arises from molecular-level interactions. CMBE, an official journal of the Biomedical Engineering Society, publishes original research and review papers in the following seven general areas: Molecular: DNA-protein/RNA-protein interactions, protein folding and function, protein-protein and receptor-ligand interactions, lipids, polysaccharides, molecular motors, and the biophysics of macromolecules that function as therapeutics or engineered matrices, for example. Cellular: Studies of how cells sense physicochemical events surrounding and within cells, and how cells transduce these events into biological responses. Specific cell processes of interest include cell growth, differentiation, migration, signal transduction, protein secretion and transport, gene expression and regulation, and cell-matrix interactions. Mechanobiology: The mechanical properties of cells and biomolecules, cellular/molecular force generation and adhesion, the response of cells to their mechanical microenvironment, and mechanotransduction in response to various physical forces such as fluid shear stress. Nanomedicine: The engineering of nanoparticles for advanced drug delivery and molecular imaging applications, with particular focus on the interaction of such particles with living cells. Also, the application of nanostructured materials to control the behavior of cells and biomolecules.