下一代测序技术（NGS）的使用对于描绘复杂人类疾病如癌症的基因突变谱系非常重要。但尽管病因候选基因变异的鉴定数量大幅增长，他们的功能仍需完全阐明，以便对患者护理有清晰的影响。单倍体人类细胞模型已成为功能基因研究的首选工具，因为它们只有一个基因组副本，因此可以显示基因变异的不遮盖表型。近年来，人类近单倍体细胞系HAP1已广泛被确认为功能遗传研究的最爱细胞系之一。它的快速周转加上只需要修改一个等位基因以表达所需的后续表型，使这个人类细胞系成为CRISPR-Cas9技术基因编辑的有价值的工具。本文回顾了HAP1细胞系模型在功能遗传研究和使用CRISPR-Cas9系统的高通量基因筛选中的最新应用。它包括其在开发新的相关疾病模型方面的应用，以进一步阐明基因功能，创造了解人类疾病基础的新途径。我们将介绍在HAP1上使用CRISPR-Cas9技术进行功能基因及NGS技术鉴定的人类单核苷酸遗传变异的有效研究方法及其对临床应用和患者护理的影响。版权所有©2023 Llargués-Sistac、Bonjoch，和Castellvi-Bel。

The use of next-generation sequencing (NGS) technologies has been instrumental in the characterization of the mutational landscape of complex human diseases like cancer. But despite the enormous rise in the identification of disease candidate genetic variants, their functionality is yet to be fully elucidated in order to have a clear implication in patient care. Haploid human cell models have become the tool of choice for functional gene studies, since they only contain one copy of the genome and can therefore show the unmasked phenotype of genetic variants. Over the past few years, the human near-haploid cell line HAP1 has widely been consolidated as one of the favorite cell line models for functional genetic studies. Its rapid turnover coupled with the fact that only one allele needs to be modified in order to express the subsequent desired phenotype has made this human cell line a valuable tool for gene editing by CRISPR-Cas9 technologies. This review examines the recent uses of the HAP1 cell line model in functional genetic studies and high-throughput genetic screens using the CRISPR-Cas9 system. It covers its use in an attempt to develop new and relevant disease models to further elucidate gene function, and create new ways to understand the genetic basis of human diseases. We will cover the advantages and potential of the use of CRISPR-Cas9 technology on HAP1 to easily and efficiently study the functional interpretation of gene function and human single-nucleotide genetic variants of unknown significance identified through NGS technologies, and its implications for changes in clinical practice and patient care.Copyright © 2023 Llargués-Sistac, Bonjoch and Castellvi-Bel.