miR-210 loss leads to widespread phenotypic and gene expression changes in human 293T cells

IntroductionHypoxia responses are critical for myriad physiological and pathological processes, such as development, tissue repair, would healing, and tumorigenesis. microRNAs (miRNAs) are a class of small non-coding RNAs that exert their functions by inhibiting the expression of their target genes,...

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Main Authors: Xiaoxiao Zhang, Zhen Meng, Chengyong Yang, Chenghao Wang, Kexin Zhang, Anxin Shi, Jingjing Guo, Yong Feng, Yan Zeng
Format: Article
Language:English
Published: Frontiers Media S.A. 2024-12-01
Series:Frontiers in Genetics
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Online Access:https://www.frontiersin.org/articles/10.3389/fgene.2024.1486252/full
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author Xiaoxiao Zhang
Zhen Meng
Chengyong Yang
Chenghao Wang
Kexin Zhang
Anxin Shi
Jingjing Guo
Yong Feng
Yan Zeng
author_facet Xiaoxiao Zhang
Zhen Meng
Chengyong Yang
Chenghao Wang
Kexin Zhang
Anxin Shi
Jingjing Guo
Yong Feng
Yan Zeng
author_sort Xiaoxiao Zhang
collection DOAJ
description IntroductionHypoxia responses are critical for myriad physiological and pathological processes, such as development, tissue repair, would healing, and tumorigenesis. microRNAs (miRNAs) are a class of small non-coding RNAs that exert their functions by inhibiting the expression of their target genes, and miR-210 is the miRNA universally and most conspicuously upregulated by hypoxia in mammalian systems. For its relationship to hypoxia, miR-210 has been studied extensively, yet no consensus exists on the roles and mechanisms of miR-210 in human physiological processes or diseases, and we know little about genuine miR-210 target genes in humans.MethodsTo better investigate the functions and mechanisms of human miR-210, therefore, we derived the human miR-210 gene knockout (KO) 293T cell lines using the CRISPR/Cas9 technology. We then examined the cellular phenotypes and gene expression profiles of 293T cells under normoxia and hypoxia conditions.Results and DiscussionWe found that the loss of miR-210 altered a variety of cellular phenotypes including proliferation and apoptosis. Subsequent global gene expression analyses identified plausible mechanisms underlying these phenotypic changes in 293T cells. In particular, we showed that miR-210 might target the expression of BNIP3L as a potential mechanism to suppress apoptosis. Surprisingly, the mRNA levels of most previously reported miR-210 target genes were not induced upon miR-210 KO, suggesting a need to reexamining and studying human miR-210 functions directly and comprehensively. Thus, our work established a human cellular system and opportunity to unravel the complexity of the regulatory networks by miR-210.
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spelling doaj-art-49ccb60bf3a94af78e146ed15286185d2025-08-20T02:38:06ZengFrontiers Media S.A.Frontiers in Genetics1664-80212024-12-011510.3389/fgene.2024.14862521486252miR-210 loss leads to widespread phenotypic and gene expression changes in human 293T cellsXiaoxiao Zhang0Zhen Meng1Chengyong Yang2Chenghao Wang3Kexin Zhang4Anxin Shi5Jingjing Guo6Yong Feng7Yan Zeng8Department of Zoology, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, ChinaDepartment of Zoology, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, ChinaDepartment of Zoology, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, ChinaDepartment of Zoology, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, ChinaDepartment of Zoology, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, ChinaDepartment of Zoology, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, ChinaCentre in Artificial Intelligence Driven Drug Discovery, Faculty of Applied Sciences, Macao Polytechnic University, Macao, ChinaInstitute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan, Hubei, ChinaDepartment of Zoology, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, ChinaIntroductionHypoxia responses are critical for myriad physiological and pathological processes, such as development, tissue repair, would healing, and tumorigenesis. microRNAs (miRNAs) are a class of small non-coding RNAs that exert their functions by inhibiting the expression of their target genes, and miR-210 is the miRNA universally and most conspicuously upregulated by hypoxia in mammalian systems. For its relationship to hypoxia, miR-210 has been studied extensively, yet no consensus exists on the roles and mechanisms of miR-210 in human physiological processes or diseases, and we know little about genuine miR-210 target genes in humans.MethodsTo better investigate the functions and mechanisms of human miR-210, therefore, we derived the human miR-210 gene knockout (KO) 293T cell lines using the CRISPR/Cas9 technology. We then examined the cellular phenotypes and gene expression profiles of 293T cells under normoxia and hypoxia conditions.Results and DiscussionWe found that the loss of miR-210 altered a variety of cellular phenotypes including proliferation and apoptosis. Subsequent global gene expression analyses identified plausible mechanisms underlying these phenotypic changes in 293T cells. In particular, we showed that miR-210 might target the expression of BNIP3L as a potential mechanism to suppress apoptosis. Surprisingly, the mRNA levels of most previously reported miR-210 target genes were not induced upon miR-210 KO, suggesting a need to reexamining and studying human miR-210 functions directly and comprehensively. Thus, our work established a human cellular system and opportunity to unravel the complexity of the regulatory networks by miR-210.https://www.frontiersin.org/articles/10.3389/fgene.2024.1486252/fullmiR-210CRISPR/Cas9target geneapoptosisBnip3L
spellingShingle Xiaoxiao Zhang
Zhen Meng
Chengyong Yang
Chenghao Wang
Kexin Zhang
Anxin Shi
Jingjing Guo
Yong Feng
Yan Zeng
miR-210 loss leads to widespread phenotypic and gene expression changes in human 293T cells
Frontiers in Genetics
miR-210
CRISPR/Cas9
target gene
apoptosis
Bnip3L
title miR-210 loss leads to widespread phenotypic and gene expression changes in human 293T cells
title_full miR-210 loss leads to widespread phenotypic and gene expression changes in human 293T cells
title_fullStr miR-210 loss leads to widespread phenotypic and gene expression changes in human 293T cells
title_full_unstemmed miR-210 loss leads to widespread phenotypic and gene expression changes in human 293T cells
title_short miR-210 loss leads to widespread phenotypic and gene expression changes in human 293T cells
title_sort mir 210 loss leads to widespread phenotypic and gene expression changes in human 293t cells
topic miR-210
CRISPR/Cas9
target gene
apoptosis
Bnip3L
url https://www.frontiersin.org/articles/10.3389/fgene.2024.1486252/full
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AT zhenmeng mir210lossleadstowidespreadphenotypicandgeneexpressionchangesinhuman293tcells
AT chengyongyang mir210lossleadstowidespreadphenotypicandgeneexpressionchangesinhuman293tcells
AT chenghaowang mir210lossleadstowidespreadphenotypicandgeneexpressionchangesinhuman293tcells
AT kexinzhang mir210lossleadstowidespreadphenotypicandgeneexpressionchangesinhuman293tcells
AT anxinshi mir210lossleadstowidespreadphenotypicandgeneexpressionchangesinhuman293tcells
AT jingjingguo mir210lossleadstowidespreadphenotypicandgeneexpressionchangesinhuman293tcells
AT yongfeng mir210lossleadstowidespreadphenotypicandgeneexpressionchangesinhuman293tcells
AT yanzeng mir210lossleadstowidespreadphenotypicandgeneexpressionchangesinhuman293tcells