Round spermatid injection (ROSI) technique holds great promise for clinical treatment of a proportion of infertile men. However, the compromised developmental potential of ROSI embryos largely limits the clinical application, and the mechanisms are not fully understood. Here, we describe the transcriptome, chromatin accessibility, and DNA methylation landscapes of mouse ROSI embryos derived from early-stage round spermatids using a single-cell multiomics sequencing approach. By interrogating these data, we identify the reprogramming defects in ROSI embryos at the pronuclear stages, which are mainly associated with the misexpression of a cohort of minor zygotic genome activation genes. We screen a small compound, A366, that can significantly increase the developmental potential of ROSI embryos, in which A366 can partially overcome the reprogramming defects by amending the epigenetic and transcriptomic states. Collectively, our study uncovers the reprogramming defects in ROSI embryos for understanding the mechanisms underlying compromised developmental potential and offers an avenue for ROSI technique optimization.

中文翻译：

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单细胞多组学测序揭示了圆形精子细胞注射产生的胚胎重编程缺陷

圆形精子细胞注射 (ROSI) 技术为部分不育男性的临床治疗提供了广阔的前景。然而，ROSI胚胎发育潜力受损在很大程度上限制了临床应用，其机制尚不完全清楚。在这里，我们使用单细胞多组学测序方法描述了源自早期圆形精子细胞的小鼠 ROSI 胚胎的转录组、染色质可及性和 DNA 甲基化景观。通过询问这些数据，我们确定了 ROSI 胚胎在原核阶段的重编程缺陷，这主要与一组次要合子基因组激活基因的错误表达有关。我们筛选了一种小化合物 A366，它可以显着提高 ROSI 胚胎的发育潜力，其中A366可以通过修改表观遗传和转录组状态来部分克服重编程缺陷。总的来说，我们的研究揭示了 ROSI 胚胎中的重编程缺陷，以了解发育潜力受损的机制，并为 ROSI 技术优化提供了途径。