Single-cell CAS-seq reveals a class of short PIWI-interacting RNAs in human oocytes

Small RNAs have important functions. However, small RNAs in primate oocytes remain unexplored. Herein, we develop CAS-seq, a single-cell small RNA sequencing method, and profiled the small RNAs in human oocytes and embryos. We discover a class of ~20-nt small RNAs that are predominantly expressed in human and monkey oocytes, but not in mouse oocytes. They are specifically associated with HIWI3 (PIWIL3), whereas significantly shorter than the commonly known PIWI-interacting RNAs (piRNAs), designated as oocyte short piRNAs (os-piRNAs). Notably, the os-piRNAs in human oocytes lack 2'-O-methylation at the 3' end, a hallmark of the classic piRNAs. In addition, the os-piRNAs have a strong 1U/10A bias and are enriched on the antisense strands of recently evolved transposable elements (TEs), indicating the potential function of silencing TEs by cleavage. Therefore, our study has identified an oocyte-specific piRNA family with distinct features and provides valuable resources for studying small RNAs in primate oocytes. Overall design: Evaluate the reproducibility and sensitivity of CAS-seq to detect small non-coding RNAs (sncRNAs) in mouse oocyte/testis and three cell lines, including HEK293, A549 and Hela. Compare the profiles of sncRNAs in the oocyte of human, mouse and monkey, and investigate the characteristics of sncRNAs and their function and expression dynamics in human oocyte and early embryos.