Single-molecule identification of the target RNAs of different RNA binding proteins simultaneously in cells.
Single-molecule identification of the target RNAs of different RNA binding proteins simultaneously in cells.
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DOI：
10.1101/gad.349983.122
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发表时间：
2022-09-01
影响因子：
10.5
通讯作者：
Meyer, Kate D.
中科院分区：
文献类型：
--
作者：
Flamand, Mathieu N.;Ke, Ke;Tamming, Renee;Meyer, Kate D.
在这种资源/方法论中，Flamand等人。开发的Tribe-Stamp，一种单分子检测两种RNA结合蛋白的靶RNA的方法。他们的数据揭示了YTHDF蛋白的共享单分子RNA靶标，并支持YTHDF1和YTHDF3的模型不太可能促进快速mRNA衰变。
RNA结合蛋白（RBP）几乎调节mRNA加工的各个方面，并且是细胞中基因表达的重要调节剂。但是，当前用于整个转录组识别RBP目标的方法是有限的，因为它们一次仅检查单个RBP，并且没有提供有关受给定RBP绑定的单个RNA分子的信息。在这里，我们通过开发Tribe-Stamp来克服这些局限性，这是一种单分子检测两种RNA结合蛋白的靶RNA的方法。我们将部落stamp应用于细胞质M6A读取器蛋白YTHDF1，YTHDF2和YTHDF3，并发现单个mRNA分子可以在其一生中受到一个以上的YTHDF蛋白，从而为细胞中YTHDF蛋白的功能提供新的见解。 Tribe-Stamp是一种高度用途的方法，可以同时在同一细胞中同时对RBP对靶标进行单分子分析。
In this Resource/Methodology, Flamand et al. developed TRIBE-STAMP, an approach for single-molecule detection of the target RNAs of two RNA binding proteins simultaneously in cells. Their data reveal shared single-molecule RNA targets of the YTHDF proteins and support a model in which YTHDF1 and YTHDF3 are unlikely to promote rapid mRNA decay. RNA-binding proteins (RBPs) regulate nearly every aspect of mRNA processing and are important regulators of gene expression in cells. However, current methods for transcriptome-wide identification of RBP targets are limited, since they examine only a single RBP at a time and do not provide information on the individual RNA molecules that are bound by a given RBP. Here, we overcome these limitations by developing TRIBE-STAMP, an approach for single-molecule detection of the target RNAs of two RNA binding proteins simultaneously in cells. We applied TRIBE-STAMP to the cytoplasmic m6A reader proteins YTHDF1, YTHDF2, and YTHDF3 and discovered that individual mRNA molecules can be bound by more than one YTHDF protein throughout their lifetime, providing new insights into the function of YTHDF proteins in cells. TRIBE-STAMP is a highly versatile approach that enables single-molecule analysis of the targets of RBP pairs simultaneously in the same cells.
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影响因子：
64.8
作者：
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作者：
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影响因子：
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48
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通讯作者：
Giallourakis CC