دورية أكاديمية

Gene editing of the multi-copy H2A.B gene and its importance for fertility

التفاصيل البيبلوغرافية
العنوان: Gene editing of the multi-copy H2A.B gene and its importance for fertility
المؤلفون: Nur Diana Anuar, Sebastian Kurscheid, Matt Field, Lei Zhang, Edward Rebar, Philip Gregory, Thierry Buchou, Josephine Bowles, Peter Koopman, David J. Tremethick, Tatiana A. Soboleva
المصدر: Genome Biology, Vol 20, Iss 1, Pp 1-16 (2019)
بيانات النشر: BMC, 2019.
سنة النشر: 2019
المجموعة: LCC:Biology (General)
LCC:Genetics
مصطلحات موضوعية: Chromatin, Histone variants, H2A.B, RNA polymerase II, Pre-mRNA splicing, Splicing speckles, Biology (General), QH301-705.5, Genetics, QH426-470
الوصف: Abstract 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.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 1474-760X
Relation: http://link.springer.com/article/10.1186/s13059-019-1633-3; https://doaj.org/toc/1474-760X
DOI: 10.1186/s13059-019-1633-3
URL الوصول: https://doaj.org/article/1ac2215f698d4f058171c99f12cd4416
رقم الأكسشن: edsdoj.1ac2215f698d4f058171c99f12cd4416
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:1474760X
DOI:10.1186/s13059-019-1633-3