Abstract
Since human and mouse Y chromosomes contain repeated sequences, it is difficult to determine the precise sequences and analyze the function of individual Y chromosome genes. Therefore, the causes of many diseases and abnormalities related to Y chromosome genes, such as male infertility, remain unclear. In this study, to elucidate the mouse Y chromosome, we enriched the mouse Y chromosome using a fluorescence-activated cell sorter (FACS) equipped with commonly used UV and blue 488 nm lasers and read the nucleotides using the Oxford Nanopore MinION long-read sequencer. This sequencing strategy allows us to cover the whole known region as well as the potential undetermined region of the Y chromosome. FACS-based chromosome enrichment and long-read sequencing are suitable for analysis of the Y chromosome sequences and may lead to further understanding of the physiological role of Y chromosome genes.
Highlights
Male mammals have a Y chromosome, but its length and the encoded genes vary greatly among species (Hughes and Page, 2015)
To enrich the Y chromosome, monocyte/macrophage RAW264.7 cells derived from BALB/c mice were treated with Colcemid to arrest the cell cycle at the M phase, and Y chromosomes were sorted by fluorescenceactivated cell sorter (FACS) according to Hoechst33342 and PI fluorescence intensities (Figure 1A)
In this study we analyzed the mouse Y chromosome using the Oxford Nanopore MinION sequencer in combination with lasers equipped for chromosome sorting
Summary
Male mammals have a Y chromosome, but its length and the encoded genes vary greatly among species (Hughes and Page, 2015). The human genome project was completed 15 years ago (Consortium, 2004), the complete sequence of the Y chromosome has not been deciphered for several reasons, including the high degree of repeated sequences and the presence of large regions of heterochromatin. Mice are often used as a model organism, and a draft sequence of the mouse genome was published without enough Y chromosome data in 2002 (Waterston et al, 2002). Thereafter, the mouse Y chromosome was re-sequenced and reported in 2014 using a BAC library covering the mouse Y chromosome (Soh et al, 2014). The complete genomic sequence of the mouse Y chromosome remains unclear because of the ampliconic region characterized by repeated sequences, which covers 98% of the chromosome (Soh et al, 2014)
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
