The human sperm is one of the smallest cells in the body, but also one of the most important, as it serves as the entire paternal genetic contribution to a child. Investigating RNA and mutations in sperm is especially relevant for diseases such as autism spectrum disorders (ASD), which have been correlated with advanced paternal age. Historically, studies have focused on the assessment of bulk sperm, wherein millions of individual sperm are present and only high-frequency variants can be detected. Using 10× Chromium single-cell sequencing technology, we assessed the transcriptome from >65,000 single spermatozoa across six sperm donors (scSperm-RNA-seq), including two who fathered multiple children with ASD and four fathers of neurotypical children. Using RNA-seq methods for differential expression and variant analysis, we found clusters of sperm mutations in each donor that are indicative of the sperm being produced by different stem cell pools. Finally, we have shown that genetic variations can be found in single sperm.

人类精子是体内最小的细胞之一，但也是最重要的细胞之一，因为它是对孩子整个父本的遗传贡献。研究精子中的RNA和突变与自闭症谱系障碍（ASD）等疾病特别相关，这些疾病已与高龄父亲年龄相关。从历史上看，研究主要集中在对大量精子的评估上，其中存在数以百万计的单个精子，并且只能检测到高频变异。使用10x铬单细胞测序技术，我们评估了来自6个精子供体（scSperm-RNA-seq）的> 65,000个单精子的转录组，其中包括两个生有多个ASD患儿和四个生神经型患儿。使用RNA-seq方法进行差异表达和变异分析，我们在每个供体中发现了一系列的精子突变，这些突变表明不同干细胞池产生的精子。最后，我们表明可以在单个精子中发现遗传变异。