Vats Sanskriti, Kumar Jitesh, Sonah Humira, Zhang Feng, Deshmukh Rupesh
National Agri-Food Biotechnology Institute (NABI), Mohali, Punjab, India.
Regional Centre for Biotechnology, Faridabad, Haryana (NCR Delhi), India.
J Exp Bot. 2024 Sep 11;75(17):5344-5356. doi: 10.1093/jxb/erae053.
Prime editors are reverse transcriptase (RT)-based genome-editing tools that utilize double-strand break (DSB)-free mechanisms to decrease off-target editing in genomes and enhance the efficiency of targeted insertions. The multiple prime editors that have been developed within a short span of time are a testament to the potential of this technique for targeted insertions. This is mainly because of the possibility of generation of all types of mutations including deletions, insertions, transitions, and transversions. Prime editing reverses several bottlenecks of gene editing technologies that limit the biotechnological applicability to produce designer crops. This review evaluates the status and evolution of the prime editing technique in terms of the types of editors available up to prime editor 5 and twin prime editors, and considers the developments in plants in a systematic manner. The various factors affecting prime editing efficiency in plants are discussed in detail, including the effects of temperature, the prime editing guide (peg)RNA, and RT template amongst others. We discuss the current obstructions, key challenges, and available resolutions associated with the technique, and consider future directions and further improvements that are feasible to elevate the efficiency in plants.
引导编辑是基于逆转录酶(RT)的基因组编辑工具,它利用无双链断裂(DSB)机制来减少基因组中的脱靶编辑,并提高靶向插入的效率。在短时间内开发出的多种引导编辑器证明了该技术在靶向插入方面的潜力。这主要是因为有可能产生包括缺失、插入、转换和颠换在内的所有类型的突变。引导编辑克服了基因编辑技术的几个瓶颈,这些瓶颈限制了生物技术在培育定制作物方面的应用。本文综述了引导编辑技术的现状和发展,涵盖了截至引导编辑器5和双引导编辑器的可用编辑器类型,并系统地探讨了其在植物领域的进展。详细讨论了影响植物引导编辑效率的各种因素,包括温度、引导编辑向导(peg)RNA和RT模板等的影响。我们讨论了与该技术相关的当前障碍、关键挑战和可用解决方案,并考虑了未来的方向以及进一步提高植物中引导编辑效率的可行改进措施。