Genome-wide single-cell analysis of recombination activity and de novo mutation rates in human sperm

Cell. 2012 Jul 20;150(2):402-12. doi: 10.1016/j.cell.2012.06.030.

Abstract

Meiotic recombination and de novo mutation are the two main contributions toward gamete genome diversity, and many questions remain about how an individual human's genome is edited by these two processes. Here, we describe a high-throughput method for single-cell whole-genome analysis that was used to measure the genomic diversity in one individual's gamete genomes. A microfluidic system was used for highly parallel sample processing and to minimize nonspecific amplification. High-density genotyping results from 91 single cells were used to create a personal recombination map, which was consistent with population-wide data at low resolution but revealed significant differences from pedigree data at higher resolution. We used the data to test for meiotic drive and found evidence for gene conversion. High-throughput sequencing on 31 single cells was used to measure the frequency of large-scale genome instability, and deeper sequencing of eight single cells revealed de novo mutation rates with distinct characteristics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Gene Conversion
  • Genome-Wide Association Study
  • Genomic Instability
  • Humans
  • Male
  • Meiosis
  • Microfluidic Analytical Techniques
  • Mutation Rate*
  • Recombination, Genetic
  • Single-Cell Analysis*
  • Spermatozoa / cytology
  • Spermatozoa / metabolism*