Single-cell m6A mapping in vivo using picoMeRIP–seq
Yanjiao Li, Yunhao Wang, María Vera-Rodríguez, Leif C. Lindeman, Linda Ellevog Skuggen, Erik M. K. Rasmussen, Ingunn Jermstad, S. Khan, Madeleine Fosslie, Trine Skuland, Marie Indahl, Sherif Khodeer, Eva Kristine Klemsdal, Kang-Xuan Jin, Knut Tomas Dalen, Péter Fedorcsák, Gareth D. Greggains, Mads Lerdrup, Arne Klungland, Kin Fai Au, John Arne Dahl
Abstract
Abstract Current N 6 -methyladenosine (m 6 A) mapping methods need large amounts of RNA or are limited to cultured cells. Through optimized sample recovery and signal-to-noise ratio, we developed picogram-scale m 6 A RNA immunoprecipitation and sequencing (picoMeRIP–seq) for studying m 6 A in vivo in single cells and scarce cell types using standard laboratory equipment. We benchmark m 6 A mapping on titrations of poly(A) RNA and embryonic stem cells and in single zebrafish zygotes, mouse oocytes and embryos.