Analyses of long-range interactions of the NR0B1 (DAX1) promoter using chromosome conformation capture-on-chip. Homo sapiens

Heterozygous point mutations or genomic deletions of NR0B1 in Xp21.2 result in congenital adrenal hypoplasia and hypogonadotropic hypogonadism, whereas the NR0B1 locus duplications in XY individuals lead to gonadal dysgenesis and a male-to-female dosage-sensitive sex reversal. We previously reported an ~ 257 kb deletion mapping 11 kb upstream to NR0B1 in a XY female with primary amenorrhea, small immature uterus, and gonadal dysgenesis pointing to an alteration of its regulatory region. To identify the potential regulatory elements of NR0B1, we have analyzed its 2 Mb flanking regions using chromosome conformation capture-on-chip (4C) in Sertoli cells and lymphoblasts. We confirm the involvement of the previously proposed regulatory region in the control of NR0B1 expression and describe several novel potential chromatin interactions within the NR0B1 locus that may be involved in sex differentiation. Overall design: Custom designed 3x720K tiling microarrays covering 4 Mb region (chrX:28,082,100-32,087,100) flanking NR0B1 gene of interest

X染色体短臂2区1带2亚区（Xp21.2）内NR0B1基因的杂合点突变或基因组缺失，可引发先天性肾上腺发育不全与促性腺激素低下型性腺功能减退症；而XY个体中NR0B1基因座的重复，则会导致性腺发育不全，并引发剂量敏感性的男-女性反转。本课题组此前曾报道1例XY女性患者，其基因组中NR0B1基因上游11kb处存在约257kb的缺失，该患者表现为原发性闭经、幼稚型小子宫与性腺发育不全，提示NR0B1基因的调控区域存在异常。为鉴定NR0B1基因的潜在调控元件，本研究采用染色质构象捕获芯片（chromosome conformation capture-on-chip, 4C）技术，对支持细胞（Sertoli cells）与淋巴母细胞中NR0B1基因上下游2Mb的侧翼区域展开分析。本研究证实了此前提出的调控区域在NR0B1基因表达调控中的作用，并描述了NR0B1基因座内数个全新的潜在染色质互作区域，这些区域可能参与性别分化过程。实验整体设计：定制设计的3×720K瓦片式微阵列芯片，覆盖目标基因NR0B1侧翼的4Mb区间（X染色体：28082100-32087100）。