Heterochiral modifications enhance robustness and function of DNA in living human cells
Heterochiral modifications enhance robustness and function of DNA in living human cells
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DOI：
10.1002/cbic.202300755
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发表时间：
2024-02-12
期刊：
影响因子：
3.2
通讯作者：
Lakin，Matthew R.
中科院分区：
文献类型：
--
作者：
Mallette，Tracy L.;Lidke，Diane S.;Lakin，Matthew R.
随着越来越多的寡核苷酸疗法被FDA批准用于治疗和疫苗接种，寡核苷酸疗法正变得越来越重要。同样，动态DNA纳米技术是一种很有前途的技术，它可以用来感知外源输入分子或内源生物标记物，并通过程序化的级联反应来整合 环境中多个传感反应的结果。这两项技术的结合可能会对生物医学产生极大的影响，因为它可以实现智能寡核苷酸疗法，这种疗法可以自主感知并对疾病状态做出反应。然而，一个特别的挑战是，由于内源核酸酶的降解，活细胞和生物体中标准核酸成分的寿命有限。在这项工作中，我们通过加入镜像、ʟ-dna核苷酸来产生异手性“缺口”来应对这一挑战。我们使用动态DNA纳米技术来证明，这些修饰使活着的人类细胞中的寡核苷酸保持完整的时间比未修饰的链更长。为此，我们使用了顺序转染法，将多个核酸输送到活的人类细胞中，同时增强了后续相互作用实际上发生在细胞内的信心。综上所述，这项工作促进了ʟ技术的发展--寡核苷酸的核酸保护和应用于 体内的DNA电路。
Oligonucleotide therapeutics are becoming increasingly important as more are approved by the FDA, both for treatment and vaccination. Similarly, dynamic DNA nanotechnology is a promising technique that can be used to sense exogenous input molecules or endogenous biomarkers and integrate the results of multiple sensing reactionsin situvia a programmed cascade of reactions. The combination of these two technologies could be highly impactful in biomedicine by enabling smart oligonucleotide therapeutics that can autonomously sense and respond to a disease state. A particular challenge, however, is the limited lifetime of standard nucleic acid components in living cells and organisms due to degradation by endogenous nucleases. In this work, we address this challenge by incorporating mirror‐image, ʟ‐DNA nucleotides to produce heterochiral “gapmers”. We use dynamic DNA nanotechnology to show that these modifications keep the oligonucleotide intact in living human cells for longer than an unmodified strand. To this end, we used a sequential transfection protocol for delivering multiple nucleic acids into living human cells while providing enhanced confidence that subsequent interactions are actually occurring within the cells. Taken together, this work advances the state of the art of ʟ‐nucleic acid protection of oligonucleotides and DNA circuitry for applicationsin vivo.