Single-cell multiomics sequencing reveals the reprogramming defects in embryos generated by round spermatid injection

Abstract

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.

Single-Cell Transcriptome Analysis Reveals Development-Specific Networks at Distinct Synchronized Antral Follicle Sizes in Sheep Oocytes

“…To analyze the stage-specific gene expression at distinct antral follicle in sheep oocytes, the cDNA of 10 oocytes was synthesized using a reverse transcription system and amplified using the Single Cell Sequence Specific Amplification Kit (Vazyme, Nanjing, China; Cat#P621-01) according to the manufacturer's instructions [85]. qPCR was conducted with the TB Green ® Premix Ex Taq™ II (Tli RNaseH Plus) (Cat#RR820L) on a StepOnePlus Real-Time PCR System (Thermo Fisher Scientific).…”

How to improve the clinical outcome of round spermatid injection (ROSI) into the oocyte: Correction of epigenetic abnormalities

“…Scriptaid treatment (250 mM, 10 h) of mouse ROSI embryos at the pronucleus stage more enhanced blastocyst formation and delivery rates restoring gene expression and abnormal DNA methylation. 21 Wang et al 100 recently found that a compound “A366” which were selected by screening of the epigenetic modification‐related small compound library was effective on normal development and delivery of mouse ROSI embryos improving epigenetic abnormalities (300 nM, 20 h). As Hosseini and Salehi, 99 confirmed, very brief exposure of round spermatids to these compounds may be equally effective and reduce predictable risks.…”