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Evaluation of N6-methyldeoxyadenosine antibody-based genomic profiling in eukaryotes
Genome Research ( IF 7 ) Pub Date : 2023-03-01 , DOI: 10.1101/gr.276696.122
Brian M Debo 1 , Benjamin J Mallory 2 , Andrew B Stergachis 3, 4, 5
Affiliation  

Low-level DNA N6-methyldeoxyadenosine (DNA-m6A) has recently been reported across various eukaryotes. Although anti-m6A antibody–based approaches are commonly used to measure DNA-m6A levels, this approach is known to be confounded by DNA secondary structures, RNA contamination, and bacterial contamination. To evaluate for these confounding features, we introduce an approach for systematically validating the selectivity of antibody-based DNA-m6A methods and use a highly selective anti-DNA-m6A antibody to reexamine patterns of DNA-m6A in C. reinhardtii, A. thaliana, and D. melanogaster. Our findings raise caution about the use of antibody-based methods for endogenous m6A quantification and mapping in eukaryotes.

中文翻译:

基于 N6-甲基脱氧腺苷抗体的真核生物基因组分析评估

最近报道了多种真核生物中存在低水平的 DNA N 6 -甲基脱氧腺苷 (DNA-m6A)。尽管基于抗 m6A 抗体的方法通常用于测量 DNA-m6A 水平,但众所周知,这种方法会受到 DNA 二级结构、RNA 污染和细菌污染的干扰。为了评估这些混杂特征,我们引入了一种系统验证基于抗体的 DNA-m6A 方法选择性的方法,并使用高选择性抗 DNA-m6A 抗体重新检查莱茵衣藻、拟南芥中 DNA- m6A模式黑腹果蝇。我们的研究结果对使用基于抗体的方法进行真核生物内源性 m6A 定量和作图提出了谨慎的态度。
更新日期:2023-03-01
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