Awwad Dahlia A
Center of X-Ray Determination of Structure of Matter (CXDS), Helmi Institute of Biomedical Research, Zewail City of Science and Technology, Giza, Cairo, Egypt.
Biol Methods Protoc. 2019 Dec 13;4(1):bpz017. doi: 10.1093/biomethods/bpz017. eCollection 2019.
Long non-coding RNAs (lncRNAs) makeup a considerable part of the non-coding human genome and had been well-established as crucial players in an array of biological processes. In spite of their abundance and versatile roles, their functional characteristics remain largely undiscovered mainly due to the lack of suitable genetic manipulation tools. The emerging CRISPR/Cas9 technology has been widely adapted in several studies that aim to screen and identify novel lncRNAs as well as interrogate the functional properties of specific lncRNAs. However, the complexity of lncRNAs genes and the regulatory mechanisms that govern their transcription, as well as their unique functionality pose several limitations the utilization of classic CRISPR methods in lncRNAs functional studies. Here, we overview the unique characteristics of lncRNAs transcription and function and the suitability of the CRISPR toolbox for applications in functional characterization of lncRNAs. We discuss some of the novel variations to the classic CRISPR/Cas9 system that have been tailored and applied previously to study several aspects of lncRNAs functionality. Finally, we share perspectives on the potential applications of various CRISPR systems, including RNA-targeting, in the direct editing and manipulation of lncRNAs.
长链非编码RNA（lncRNAs）构成了人类非编码基因组的相当一部分，并已被确认为一系列生物学过程中的关键参与者。尽管它们数量众多且作用多样，但其功能特性在很大程度上仍未被发现，主要原因是缺乏合适的基因操作工具。新兴的CRISPR/Cas9技术已在多项研究中广泛应用，这些研究旨在筛选和鉴定新型lncRNAs以及探究特定lncRNAs的功能特性。然而，lncRNAs基因的复杂性、调控其转录的机制以及它们独特的功能，给经典CRISPR方法在lncRNAs功能研究中的应用带来了一些限制。在此，我们概述lncRNAs转录和功能的独特特征以及CRISPR工具包在lncRNAs功能表征应用中的适用性。我们讨论了一些经典CRISPR/Cas9系统的新型变体，这些变体此前已被定制并应用于研究lncRNAs功能的多个方面。最后，我们分享了对各种CRISPR系统（包括RNA靶向）在lncRNAs直接编辑和操作中的潜在应用的看法。
