Gene editing of the multi-copy H2A.B gene and its importance for fertility
| Autor: | Peter Koopman, David J. Tremethick, Tatiana A. Soboleva, Lei Zhang, Josephine Bowles, Matthew A. Field, Sebastian Kurscheid, Philip D. Gregory, Thierry Buchou, Nur Diana Anuar, Edward J. Rebar |
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| Rok vydání: | 2019 |
| Předmět: |
Male
lcsh:QH426-470 Chromosomal Proteins Non-Histone Gene Expression RNA polymerase II Histones 03 medical and health sciences Pre-mRNA splicing 0302 clinical medicine Transcription Activator-Like Effector Nucleases Gene expression Animals Gene family lcsh:QH301-705.5 Gene Infertility Male 030304 developmental biology Gene Editing Mice Knockout Histone variants Genetics 0303 health sciences Base Sequence biology Research Splicing speckles Spermatozoa Chromatin lcsh:Genetics TALENs Fertility Histone lcsh:Biology (General) Mutation RNA splicing Knockout mouse biology.protein Female H2A.B 030217 neurology & neurosurgery Genome editing |
| Zdroj: | Genome Biology Genome Biology, Vol 20, Iss 1, Pp 1-16 (2019) |
| ISSN: | 1474-760X |
| DOI: | 10.1186/s13059-019-1633-3 |
| Popis: | Background Altering the biochemical makeup of chromatin by the incorporation of histone variants during development represents a key mechanism in regulating gene expression. The histone variant H2A.B, H2A.B.3 in mice, appeared late in evolution and is most highly expressed in the testis. In the mouse, it is encoded by three different genes. H2A.B expression is spatially and temporally regulated during spermatogenesis being most highly expressed in the haploid round spermatid stage. Active genes gain H2A.B where it directly interacts with polymerase II and RNA processing factors within splicing speckles. However, the importance of H2A.B for gene expression and fertility are unknown. Results Here, we report the first mouse knockout of this histone variant and its effects on fertility, nuclear organization, and gene expression. In view of the controversy related to the generation of off-target mutations by gene editing approaches, we test the specificity of TALENs by disrupting the H2A.B multi-copy gene family using only one pair of TALENs. We show that TALENs do display a high level of specificity since no off-target mutations are detected by bioinformatics analyses of exome sequences obtained from three consecutive generations of knockout mice and by Sanger DNA sequencing. Male H2A.B.3 knockout mice are subfertile and display an increase in the proportion of abnormal sperm and clogged seminiferous tubules. Significantly, a loss of proper RNA Pol II targeting to distinct transcription–splicing territories and changes to pre-mRNA splicing are observed. Conclusion We have produced the first H2A.B knockout mouse using the TALEN approach. Electronic supplementary material The online version of this article (10.1186/s13059-019-1633-3) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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