Advances in molecular technologies over the past few decades, such as high-throughput DNA marker genotyping, have provided more powerful plant breeding approaches, including marker-assisted selection and genomic selection. At the same time, massive investments in plant genetics and genomics, led by whole genome sequencing, have led to greater knowledge of genes and genetic pathways across plant genomes. However, there remains a gap between approaches focused on forward genetics, which start with a phenotype to map a mutant locus or QTL with the goal of cloning the causal gene, and approaches using reverse genetics, which start with large-scale sequence data and work back to the gene function. The recent establishment of efficient CRISPR-Cas-based gene editing promises to bridge this gap and provide a rapid method to functionally validate genes and alleles identified through studies of natural variation. CRISPR-Cas techniques can be used to knock out single or multiple genes, precisely modify genes through base and prime editing, and replace alleles. Moreover, technologies such as protoplast isolation, in planta transformation, and the use of developmental regulatory genes promise to enable high-throughput gene editing to accelerate crop improvement.

中文翻译：

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通过基因编辑进行功能等位基因验证以利用丰富的遗传资源进行作物改良

过去几十年分子技术的进步，例如高通量 DNA 标记基因分型，提供了更强大的植物育种方法，包括标记辅助选择和基因组选择。与此同时，以全基因组测序为主导的对植物遗传学和基因组学的大量投资，使人们对植物基因组中的基因和遗传途径有了更多的了解。然而，在专注于正向遗传学的方法（从表型开始绘制突变基因座或 QTL 以克隆因果基因）与使用反向遗传学的方法（从大规模序列数据和工作开始）之间仍然存在差距回到基因功能。最近建立的基于 CRISPR-Cas 的高效基因编辑有望弥合这一差距，并提供一种快速方法来功能验证通过研究自然变异鉴定的基因和等位基因。CRISPR-Cas技术可用于敲除单个或多个基因，通过碱基和引物编辑精确修饰基因，替换等位基因。此外，原生质体分离、植物转化和发育调控基因的使用等技术有望实现高通量基因编辑以加速作物改良。