Department of Biology, Swarthmore College, Swarthmore, PA, USA.
Department of Molecular Phamacology, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
Chromosome Res. 2020 Mar;28(1):87-110. doi: 10.1007/s10577-019-09625-x. Epub 2020 Feb 6.
The nucleus is a highly organized and dynamic environment where regulation and coordination of processes such as gene expression and DNA replication are paramount. In recent years, noncoding RNAs have emerged as key participants in the regulation of nuclear processes. There are a multitude of functional roles for long noncoding RNA (lncRNA), mediated through their ability to act as molecular scaffolds bridging interactions with proteins, chromatin, and other RNA molecules within the nuclear environment. In this review, we discuss the diversity of techniques that have been developed to probe the function of nuclear lncRNAs, along with the ways in which those techniques have revealed insights into their mechanisms of action. Foundational observations into lncRNA function have been gleaned from molecular cytology-based, single-cell approaches to illuminate both the localization and abundance of lncRNAs in addition to their potential binding partners. Biochemical, extraction-based approaches have revealed the molecular contacts between lncRNAs and other molecules within the nuclear environment and how those interactions may contribute to nuclear organization and regulation. Using examples of well-studied nuclear lncRNAs, we demonstrate that the emerging functions of individual lncRNAs have been most clearly deduced from combined cytology and biochemical approaches tailored to study specific lncRNAs. As more functional nuclear lncRNAs continue to emerge, the development of additional technologies to study their interactions and mechanisms of action promise to continually expand our understanding of nuclear organization, chromosome architecture, genome regulation, and disease states.
核是一个高度组织化和动态的环境，其中基因表达和 DNA 复制等过程的调节和协调至关重要。近年来，非编码 RNA 已成为调节核过程的关键参与者。长链非编码 RNA (lncRNA) 具有多种功能作用，其通过作为分子支架的能力，与核环境中的蛋白质、染色质和其他 RNA 分子相互作用。在这篇综述中，我们讨论了已经开发出的多种技术，用于研究核 lncRNA 的功能，以及这些技术如何揭示它们的作用机制。从基于分子细胞学的单细胞方法中获得的 lncRNA 功能的基础观察结果，不仅阐明了 lncRNA 的定位和丰度，还阐明了它们潜在的结合伙伴。基于生化、提取的方法揭示了核环境中 lncRNA 与其他分子之间的分子接触，以及这些相互作用如何有助于核组织和调节。通过研究一些研究充分的核 lncRNA 的例子，我们证明了从专门研究特定 lncRNA 的细胞学和生化方法的结合中，可以最清楚地推断出个别 lncRNA 的新兴功能。随着更多功能核 lncRNA 的不断出现，开发更多用于研究它们相互作用和作用机制的技术，有望不断扩展我们对核组织、染色体结构、基因组调节和疾病状态的理解。
