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Can genetic engineering-based methods for gene function identification be eclipsed by genome editing in plants? A comparison of methodologies
Molecular Genetics and Genomics ( IF 2.3 ) Pub Date : 2021-03-09 , DOI: 10.1007/s00438-021-01769-y
P. P. Amritha , Jasmine M. Shah

Finding and explaining the functions of genes in plants have promising applications in crop improvement and bioprospecting and hence, it is important to compare various techniques available for gene function identification in plants. Today, the most popular technology among researchers to identify the functions of genes is the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9)-based genome editing method. But by no means can we say that CRISPR/Cas9 is the go-to method for all purposes. It comes with its own baggage. Researchers will agree and have lived through at least seven more technologies deployed to find the functions of genes, which come under three umbrellas: 1. genetic engineering, 2. transient expression, and 3. chemical/physical mutagenesis. Each of the methods evolved when the previous one ran into an insurmountable problem. In this review, we compare the eight technologies against one another on 14 parameters. This review lays bare the pros and cons, and similarities and dissimilarities of various methods. Every method comes with its advantages and disadvantages. For example, the CRISPR/Cas9-based genome editing is an excellent method for modifying gene sequences, creating allelic versions of genes, thereby aiding the understanding of gene function. But it comes with the baggage of unwanted or off-target mutations. Then, we have methods based on random or targeted knockout of the gene, knockdown, and overexpression of the gene. Targeted disruption of genes is required for complete knockout of gene function, which may not be accomplished by editing. We have also discussed the strategies to overcome the shortcomings of the targeted gene-knockout and the CRISPR/Cas9-based methods. This review serves as a comprehensive guide towards the understanding and comparison of various technologies available for gene function identification in plants and hence, it will find application for crop improvement and bioprospecting related research.



中文翻译:

植物中的基因组编辑能否使基于基因工程的基因功能识别方法黯然失色?方法论比较

发现和解释植物中基因的功能在作物改良和生物勘探中具有广阔的应用前景,因此,比较各种可用于植物中基因功能鉴定的技术非常重要。如今,研究人员中鉴定基因功能的最流行技术是基于规则间隔的簇状短回文重复序列(CRISPR)/ CRISPR相关蛋白9(Cas9)的基因组编辑方法。但是我们绝对不能说CRISPR / Cas9是实现所有目的的首选方法。它带有自己的行李。研究人员将同意并已经经历了至少七种用于发现基因功能的技术,这些技术可以归为三大类:1.基因工程,2。瞬时表达和3.化学/物理诱变。当前一个方法遇到无法解决的问题时,每种方法都在发展。在这篇综述中,我们在14个参数上比较了这8种技术。这篇评论揭示了各种方法的利弊,以及各种方法的异同。每种方法都有其优点和缺点。例如,基于CRISPR / Cas9的基因组编辑是修改基因序列,创建基因的等位基因版本,从而有助于对基因功能的理解的极佳方法。但是它带来了不必要的或偏离目标的突变。然后,我们有了基于基因的随机或有针对性的敲除,敲除和基因过表达的方法。基因敲除是完全敲除基因功能所必需的,这可能无法通过编辑来完成。我们还讨论了克服靶向基因敲除和基于CRISPR / Cas9的方法的缺点的策略。这篇综述作为理解和比较可用于植物基因功能鉴定的各种技术的综合指南,因此,它将在作物改良和生物勘探相关研究中得到应用。

更新日期:2021-03-10
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