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全基因组CRISPR/Cas9筛选鉴定干细胞衰老的调控基因。

A Genome-Wide CRISPR/Cas9 Screen Identifies Regulatory Genes for Stem Cell Aging.

作者信息

Su Peng, Miao Yi-Liang

机构信息

Institute of Stem Cell and Regenerative Biology, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, China.

出版信息

Methods Mol Biol. 2025;2960:159-170. doi: 10.1007/7651_2024_566.

Abstract

Aging is a ubiquitous biological phenomenon, characterized by a gradual decline in physiological functions and an increased risk of various diseases. Although it is known that aging involves extensive changes in gene expression and disruptions in cellular metabolism, the molecular mechanisms underlying these processes remain incompletely understood. The CRISPR/Cas9 technology provides an efficient method for gene editing. In recent years, this technique has been successfully applied in various cellular and animal models to identify key genes involved in biological processes such as cancer and genetic diseases, which makes it possible to screen genes that affect cell senescence in the whole genome. Here, we describe a method that involves differentiating embryonic stem cells into mesenchymal progenitor cells and employing CRISPR/Cas9 for genome-wide functional screening to identify genes that regulate aging. Further analysis of the functions and regulatory mechanisms of these genes may provide new targets and strategies for anti-aging research and stem cell therapy.

摘要

衰老 是一种普遍存在的生物学现象,其特征是生理功能逐渐衰退以及患各种疾病的风险增加。尽管已知衰老涉及基因表达的广泛变化和细胞代谢的紊乱,但这些过程背后的分子机制仍未完全了解。CRISPR/Cas9技术为基因编辑提供了一种有效的方法。近年来,该技术已成功应用于各种细胞和动物模型,以鉴定参与癌症和遗传疾病等生物学过程的关键基因,这使得在全基因组范围内筛选影响细胞衰老的基因成为可能。在此,我们描述了一种方法,该方法包括将胚胎干细胞分化为间充质祖细胞,并采用CRISPR/Cas9进行全基因组功能筛选,以鉴定调节衰老的基因。对这些基因的功能和调控机制进行进一步分析,可能为抗衰老研究和干细胞治疗提供新的靶点和策略。

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