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定量基础分辨率测序技术用于绘制哺乳动物mRNA中伪乌里丁的映射

Li-Sheng Zhang¹, Qing Dai², Chuan He²

¹Division of Life Science, The Hong Kong University of Science and Technology (HKUST), Hong Kong, SAR, P.R. China; Department of Chemistry, The Hong Kong University of Science and Technology (HKUST), Hong Kong, SAR, P.R. China.

Methods in enzymology

|November 4, 2023

概括

研究人员开发了BID-seq,这是一种新的方法,用于在信使RNA (mRNA) 转录组中映射伪尿素 (Ψ). 这种技术量化了 Ψ 修饰固体几何学,使其能够更深入地了解其生物学作用.

科学领域:

分子生物学分子生物学
表观遗传学在表观遗传学中,表观遗传学是指表观遗传学.
在RNA生物学,RNA生物学.

背景情况:

转录后RNA修饰在所有生命形式中都至关重要.
伪氨基酸 (Ψ) 是哺乳动物中丰富的RNA修饰物,调节各种RNA物种,如rRNA,tRNA和snRNA.
由于缺乏高效的转录组全方位映射方法,调查mRNA伪尿素 (Ψ) 功能是具有挑战性的.

研究的目的:

开发一种定量方法,用于在信使RNA (mRNA) 中对伪尿素 (Ψ) 的转录组范围的映射.
为了使mRNA伪尿素 (Ψ) 在各种生物过程中的功能能够得到调查.

主要方法:

开发了双硫酸盐诱导的删除序列 (BID-seq),是一种新的化学反应.
优化基于二硫酸盐的反应,可选择性地将伪乌里丁 (Ψ) 转化为 Ψ-BS 添加物,而无需细胞因子去胺.
在逆转录过程中利用了删除签名来量化每个位点的 Ψ 修饰静态度.

主要成果:

BID-seq有效地绘制成千上万个mRNA Ψ位点的地图,并从最小的RNA输入 (10-20 ng多A+RNA) 获得石基度信息.
在哺乳动物mRNA中确定了 Ψ 的共识动机,并为 Ψ 沉积分配了特定的"编写器"蛋白.
证实了哺乳动物mRNA停止符号中存在 Ψ 的存在.

结论:

BID-seq提供了一种强大的工具,用于对mRNA伪尿素 (Ψ) 的定量,转录组范围的映射.
这种方法有助于研究 Ψ 固体测量及其在基因调节中的作用.
BID-seq为未来研究mRNA Ψ在各种生物环境中的功能意义奠定了基础.

关键词:

化学生物学是化学生物学.表观遗传学在表观遗传学中,表观遗传学是指表观遗传学.这是下一代测序.伪氨酸是一种伪氨酸.基因组RNA的修饰是RNA的修饰.

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