Saha Dipnarayan, Panda Alok Kumar, Datta Subhojit
Biotechnology Unit, ICAR-Central Research Institute for Jute and Allied Fibres, Barrackpore, Kolkata, West Bengal, 700121, India.
Heliyon. 2024 Dec 14;11(1):e41135. doi: 10.1016/j.heliyon.2024.e41135. eCollection 2025 Jan 15.
Recent advances in genome editing tools and CRISPR-Cas technologies have enabled plant genome engineering reach new heights. The current regulatory exemptions for certain categories of genome edited products, such as those derived from SDN-1 and SDN-2, which are free of any transgene, have significantly accelerated genome editing research in a number of agricultural crop plants in different countries. Although CRISPR-Cas technology is becoming increasingly popular, it is still important to carefully consider a number of factors before planning and carrying conducting CRISPR-Cas studies. To attempt genome editing in a plant, a high-quality genome sequence and a repeatable tissue culture protocol for in vitro regeneration are essential. One of the most important steps in plant genome editing is the designing of a CRISPR construct, which involves selecting the appropriate Cas protein, sgRNA sequence, and appropriate regulatory sequence to trigger expression. Computational tools and algorithms play a crucial role in construct design and gRNA selection to minimize off-target effects and also to optimize their delivery techniques. Researchers may need to select appropriate software tools capable of analyzing post-editing detection of mutation events and other DNA sequence abnormalities to identify off-target effects. To fully fulfill the potential of plant genome editing, continued advances in computational biology are essential to meet the challenges it faces today.
基因组编辑工具和CRISPR-Cas技术的最新进展使植物基因组工程达到了新高度。目前对某些类别的基因组编辑产品的监管豁免,例如那些源自SDN-1和SDN-2且不含任何转基因的产品,显著加速了不同国家许多农作物的基因组编辑研究。尽管CRISPR-Cas技术越来越受欢迎,但在规划和开展CRISPR-Cas研究之前,仔细考虑一些因素仍然很重要。要在植物中尝试基因组编辑,高质量的基因组序列和用于体外再生的可重复组织培养方案至关重要。植物基因组编辑中最重要的步骤之一是设计CRISPR构建体,这涉及选择合适的Cas蛋白、sgRNA序列和合适的调控序列以触发表达。计算工具和算法在构建体设计和gRNA选择中起着关键作用,以尽量减少脱靶效应并优化其递送技术。研究人员可能需要选择能够分析编辑后突变事件检测和其他DNA序列异常的合适软件工具,以识别脱靶效应。为了充分发挥植物基因组编辑的潜力,计算生物学的持续进步对于应对当今面临的挑战至关重要。