Whole genome sequencing approach for the detection of rare protein-altering variants associated with familial male infertility.

Background: Whole-genome sequencing (WGS) is an efficient, accurate, and high-throughput technique used for the discovery of exonic as well as non-exonic variants causing different genetic anomalies, improving clinical management for the disease. This study was designed to evaluate pathogenic genetic variants related to male infertility in male infertile families in remote rural areas of Pakistan. Methods: For the five families recruited, WGS was conducted on 14 patients and 6healthy controls. WGS was performed using the Illumina TruSeq DNA Nano Library Prep workflow protocol. DNA was assessed with TapeStation and sequenced using the TruSeq kit (Illumina). Bioinformatics tools were then used to identify potential causative variants, including indels and single-nucleotide variants. Results: In family 1, five homozygous nonsynonymous SNVs were identified in Conclusion: The study contributes to a systematic understanding of the problem and provides a basis for identifying causal variations in idiopathic cases of male infertility, setting targets for future therapeutic development.