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Uri Drucker, was a political candidate for the local elections in Kiryat Tivon in 2018.
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Studying the role of the non-coding genome during sex determination
A main research interest of our lab is to understand, at the molecular level, how sex is determined during embryonic development, i.e., how does an embryo develop into either male or female.
In mammals, sex determination is genetically driven with XY individuals developing as males and XX individuals developing as females. However, if the process of sex determination is impaired, patients appear as XY females or XX males (sex reverse). These are all classified as patients with Disorders of Sex Development (DSD), with a prevalence of 1: 2500-4000 newborns.
During embryonic development, the gonads initially develop as a bipotential organ and it is the activity of several key transcription factors that directs them to develop into either testes or ovaries. In the lab, we use the mouse as a model system and are interested at understanding the regulation of these key transcription factors and how these interact with each other. We also study the role of the non-coding genome in mediating the process of sex determination and DSD pathologies.
We employ cutting-edge techniques to address these questions including CRISPR/Cas9 genome editing, transgenic mice production , advanced sequencing techniques as well as microscopy and molecular biology.
Developing an in vitro model to study the gonads
In an attempt to develop an in vitro system that allows to model the mammalian gonads we use mouse Embryonic Stem Cells (ESC) and develop differentiation protocols towards gonadal cell type of the gonads. In addition, we use similar protocols with Human ESC and induced Pluripotent Stem Cells to model DSD patients in vitro. Furthermore, we perform tissue engineering to model the testis and the spermatogenesis process using these stem cell-derived somatic cells along with germ cells in a 3D culture system.
Cell fate decisions require appropriate regulation of key genes. Sox9, a direct target of SRY, is pivotal in mammalian sex determination. In vivo high-throughput chromatin accessibility techniques, transgenic assays, and genome editing revealed several novel gonadal regulatory elements in the 2-megabase gene desert upstream of Sox9. Although others are redundant, enhancer 13 (Enh13), a 557–base pair element located 565 kilobases 5′ from the transcriptional start site, is essential to initiate mouse testis development; its deletion results in XY females with Sox9 transcript levels equivalent to those in XX gonads. Our data are consistent with the time-sensitive activity of SRY and indicate Drucker a strict order of enhancer usage. Enh13 is conserved and embedded within a 32.5-kilobase region whose deletion in humans is associated with XY sex reversal, suggesting that it is also critical in humans.
The regulation of genes with important roles in embryonic development can be complex, involving multiple, often redundant enhancers, silencers, and insulators (1, 2). The genes may have a poised epigenetic state prior to their expression, and their activation or repression may involve positive or negative feed-forward loops. This complexity is likely to be amplified when the gene has functions in more than one tissue, given that the regulatory elements required for each are often interspersed and necessitate dynamic alterations in chromatin conformation (1, 2).
The developing gonads constitute an interesting system in which to explore questions of gene regulation during development (3). Most of the cell lineages are bipotential, with the ability to give rise to cell types typical of either ovaries or testes, and many genes that become associated with male or female fate begin by being expressed at equivalent, although usually low, levels in supporting cell precursors of both XX and XY gonads (4–
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In mammals, the Sry gene encodes a protein that is transiently expressed and initiates testis and subsequent male development by triggering cells of the supporting cell lineage to differentiate into Sertoli cells rather than granulosa cells typical of ovaries (7). Sox9, the main target of SRY, is critical for the differentiation of Sertoli cells and then functions along with other transcription factors, notably Sox8 and then Dmrt1, for Sertoli cell maintenance (4–6). Both gain- and loss-of-function studies in mice and humans demonstrate that Sox9 plays a key role in testis determination (8–13). Notably, humans heterozygous for null mutations develop campomelic dysplasia (CD) [Online Mendelian Inheritance in Man (OMIM) entry 114290] (11), a severe syndrome where 70% of XY patients show female development (12, 13).
Sox9 functions in many embryonic and adult cell types (14), and genetic and molecular evidence suggests that its regulatory region is spread over a gene desert of at least 2 Mb 5′ to the coding sequence (15). The only enhancer known to be relevant for expression in Sertoli cells was TES, a 3.2-kb element mapping 13 kb 5′ from the transcriptional start site, and its 1.4-kb core, TESCO (16). Targeted deletion of TES or TESCO reduced Sox9 expression levels in the early and postnatal mouse testis to about 45% of normal but did not result in XY female development (17).
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We therefore used several unbiased approaches to systematically screen for additional gonad enhancers upstream of mouse Sox9. We used deoxyribonuclease I hypersensitive site sequencing (DNaseI-seq) data obtained with embryonic day 13.5 (E13.5) and E15.5 sorted Sertoli cells (18). From 33 putative enhancers, we chose only those positive at both stages (14 enhancers) for in vivo validation by transgenic assays (Fig. 1A and fig. S1).
In parallel, we carried out ATAC-seq (assay for transposase-accessible chromatin using sequencing) on XY and XX gonads, which permitted the use of fewer sorted cells at E10.5, an early bipotential stage, and E13.5, when gonadal sex is already determined (figs. S1 and S2 and methods). Most putative enhancers discovered by DNaseI-seq were evident in the E13.5 XY ATAC-seq data; however, we used this assay to include two more putative enhancers in the in vivo screen: enhancer 1 (Enh1) and Enh14 (Fig. 1A and fig. S1). Chromatin immunoprecipitation sequencing (ChIP-seq) was also performed for H3K27ac, a histone modification that marks active enhancers (fig. S1).
A database of more than 56,000 persons who fled to neutral Switzerland during World
War II is available at http://etat.geneve.ch/dt/archives/a_votre_service-liste_refugies1700.html. The complete list is presented alphabetically. Information about each
individual includes name, date of birth and nationality. The site is in French. Use Google
translate to convert the descriptive information in your native language.
Descriptive Catalogue of Hebrew MSS. of the Montefiore Library
The Jewish Quarterly Review
The Jewish Quarterly Review
Vol. 14, No. 2 (Jan., 1902), pp. 379-412 (34 pages)
Published by: University of Pennsylvania Press