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Beyond classic editing: innovative CRISPR approaches for functional studies of long non-coding RNA

机译:超越经典编辑:长期非编码RNA的功能研究的创新克隆方法

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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.
机译:长期非编码RNA(LNCRNA)构成了非编码人类基因组的相当大部分,并且已经在一系列生物过程中作为关键球员良好。尽管他们丰富和多功能的角色,但它们的功能特征在很大程度上仍然是由于缺乏合适的遗传操作工具。新兴的CRISPR / CAS9技术在旨在筛选和识别新型LNCRNA以及询问特定LNCRNA的功能性质的几项研究中,该技术已被广泛适应。然而,LNCRNA基因的复杂性和治理转录的监管机制以及它们独特的功能造成了几个限制,利用经典CISPR方法在LNCRNA功能研究中的利用。在这里,我们概述了LNCRNA转录和功能的独特特征以及CRISPR工具箱在LNCRNA的功能表征中的应用程序的适用性。我们讨论了对经典的CRAP / CAS9系统进行了一些新的变化,这些变化是在以前测量的,以研究LNCRNA功能的几个方面。最后,我们在直接编辑和操纵LNCRNA的直接编辑和操纵中,我们分享了各种CRISP系统的潜在应用,包括RNA靶向的潜在应用。

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