Agricultural Biotechnology Division, National Institute for Biotechnology & Genetic Engineering (NIBGE), Faisalabad, Pakistan.
Pakistan Agriculture Research Council, Islamabad, Pakistan.
J Cell Physiol. 2020 Feb;235(2):666-682. doi: 10.1002/jcp.29052. Epub 2019 Jul 10.
Clustered regularly interspaced short palindromic repeats (CRISPR)-based genome editing, derived from prokaryotic immunity system, is rapidly emerging as an alternative platform for introducing targeted alterations in genomes. The CRISPR-based tools have been deployed for several other applications including gene expression studies, detection of mutation patterns in genomes, epigenetic regulation, chromatin imaging, etc. Unlike the traditional genetic engineering approaches, it is simple, cost-effective, and highly specific in inducing genetic variations. Despite its popularity, the technology has limitations such as off-targets, low mutagenesis efficiency, and its dependency on in-vitro regeneration protocols for the recovery of stable plant lines. Several other issues such as persisted CRISPR activity in subsequent generations, the potential for transferring to its wild type population, the risk of reversion of edited version to its original phenotype particularly in cross-pollinated plant species when released into the environment and the scarcity of validated targets have been overlooked. This article briefly highlights these undermined aspects, which may challenge the wider applications of this platform for improving crop genetics.
基于成簇规律间隔短回文重复序列（CRISPR）的基因组编辑技术源自原核生物的免疫防御系统，它迅速成为在基因组中引入靶向改变的替代平台。基于 CRISPR 的工具已被用于包括基因表达研究、基因组中突变模式的检测、表观遗传调控、染色质成像等在内的其他几个应用。与传统的遗传工程方法不同，它在诱导遗传变异方面具有简单、经济高效和高度特异性的特点。尽管这项技术很受欢迎，但它也存在一些局限性，例如脱靶效应、低突变效率以及对体外再生方案的依赖，以恢复稳定的植物株系。还有其他一些问题，如 CRISPR 在后续世代中的持续活性、转移到其野生型种群的可能性、编辑版本回复到其原始表型的风险，特别是在交叉授粉植物物种中，当它们被释放到环境中时，以及有效的验证靶点的缺乏，这些都被忽视了。本文简要地强调了这些被忽视的方面，这些方面可能会挑战该平台在改善作物遗传方面的更广泛应用。
