Allemailem Khaled S, Rahmani Arshad Husain, Almansour Nahlah Makki, Aldakheel Fahad M, Albalawi Ghadah Mohammad, Albalawi Ghadeer Mohammed, Khan Amjad Ali
Department of Medical Laboratories, College of Applied Medical Sciences, Qassim University, Buraydah 51452, Saudi Arabia.
Department of Biology, College of Science, University of Hafr Al Batin, Hafr Al Batin 31991, Saudi Arabia.
Int J Oncol. 2025 May;66(5). doi: 10.3892/ijo.2025.5748. Epub 2025 May 9.
For centuries, a competitive evolutionary race between prokaryotes and related phages or other mobile genetic elements has led to the diversification of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR‑associated sequence (Cas) genome‑editing systems. Among the different CRISPR/Cas systems, the CRISPR/Cas9 system has been widely studied for its precise DNA manipulation; however, due to certain limitations of direct DNA targeting, off‑target effects and delivery challenges, researchers are looking to perform transient knockdown of gene expression by targeting RNA. In this context, the more recently discovered type VI CRISPR/Cas13 system, a programmable single‑subunit RNA‑guided endonuclease system that has the capacity to target and edit any RNA sequence of interest, has emerged as a powerful platform to modulate gene expression outcomes. All the Cas13 effectors known so far possess two distinct ribonuclease activities. Pre‑CRISPR RNA processing is performed by one RNase activity, whereas the two higher eukaryotes and prokaryotes nucleotide‑binding domains provide the other RNase activity required for target RNA degradation. Recent innovative applications of the type VI CRISPR/Cas13 system in nucleic acid detection, viral interference, transcriptome engineering and RNA imaging hold great promise for disease management. This genome editing system can also be employed by the Specific High Sensitivity Enzymatic Reporter Unlocking platform to identify any tumor DNA. The discovery of this system has added a new dimension to targeting, tracking and editing circulating microRNA/RNA/DNA/cancer proteins for the management of cancer. However, there is still a lack of thorough understanding of the mechanisms underlying some of their functions. The present review summarizes the recent updates on the type VI CRISPR/Cas system in terms of its structural and mechanistic properties and some novel applications of this genome‑editing tool in cancer management. However, some issues, such as collateral degradation of bystander RNA, impose major limitations on its application. Furthermore, additional challenges and future prospects for this genome editing system are described in the present review.
几个世纪以来，原核生物与相关噬菌体或其他可移动遗传元件之间的竞争性进化竞赛导致了成簇规律间隔短回文重复序列（CRISPR）和CRISPR相关序列（Cas）基因组编辑系统的多样化。在不同的CRISPR/Cas系统中，CRISPR/Cas9系统因其精确的DNA操作而得到广泛研究；然而，由于直接DNA靶向的某些局限性、脱靶效应和递送挑战，研究人员正在寻求通过靶向RNA来实现基因表达的瞬时敲低。在这种背景下，最近发现的VI型CRISPR/Cas13系统，一种可编程的单亚基RNA引导的内切核酸酶系统，能够靶向和编辑任何感兴趣的RNA序列，已成为调节基因表达结果的强大平台。目前已知的所有Cas13效应器都具有两种不同的核糖核酸酶活性。前CRISPR RNA加工由一种核糖核酸酶活性执行，而两个高等真核生物和原核生物核苷酸结合结构域提供靶RNA降解所需的另一种核糖核酸酶活性。VI型CRISPR/Cas13系统在核酸检测、病毒干扰、转录组工程和RNA成像方面的最新创新应用对疾病管理具有巨大潜力。这种基因组编辑系统还可被特异性高灵敏度酶促报告分子解锁平台用于识别任何肿瘤DNA。该系统的发现为靶向、追踪和编辑循环中的微小RNA/RNA/DNA/癌症蛋白以管理癌症增添了新的维度。然而，对于其某些功能背后的机制仍缺乏深入了解。本综述总结了VI型CRISPR/Cas系统在结构和机制特性方面的最新进展，以及这种基因组编辑工具在癌症管理中的一些新应用。然而，一些问题，如旁观者RNA的附带降解，对其应用造成了重大限制。此外，本综述还描述了这种基因组编辑系统的其他挑战和未来前景。
