Identifying the RP10 gene causing retinitis pigmentosa

描述(申请人提供)：该项目的目的是鉴定，即“克隆”导致常染色体显性遗传性视网膜色素变性(Adrp)的RP10基因。在初步研究中，我们通过连锁定位将RP10基因定位于人类染色体7q31，定位于4个不相关的大的adrp家族。含有RP10基因的区域由D7S2471和RP-STR12标记组成，全长3.3 MBP。通过结合公开的(GenBank)和私有的(Celera)人类基因组序列，我们已经组装了该区域99%的完整序列。同样，Celera数据库包含99%的同音鼠标序列。通过计算分析和实验室方法，我们已经在该区域确定了38个独立的表达序列，其中10个是视网膜表达的，因此可能是疾病基因候选。我们建议在这些发现和资源的基础上完成RP10基因的克隆任务。 提出的目标是1)减少RP10连锁区域，2)完成该区域的物理和转录图谱，3)测试RP10家族的候选基因。目标1的方法包括增强连锁分析和SNP单倍型测试，以完善断裂点；以及从患者那里生产单倍体细胞系，以便于测试。目标2的方法包括进一步的计算分析，比较老鼠和人类的基因组；以及根据转录谱和功能确定候选基因的优先顺序。目标3的方法包括使用基因敲除小鼠检测该区域视网膜和光感受器表达的基因；候选基因筛选；以及在无关的ADRP患者中确认阳性结果。这些研究将与南非国家生物信息学研究所和都柏林三一学院的眼遗传学单位合作完成。 RP10基因的突变可能导致至少10%的ADRP，进而影响成千上万的美国人。RP10基因的鉴定将通过诊断和咨询为受影响的个人提供直接好处。此外，这也是设计基因特异性疗法和确定修饰因素的必要的第一步。更广泛地说，对导致视网膜色素变性的基因的鉴定揭示了视网膜中新的功能通路，因此克隆RP10基因可能有助于更好地理解正常的视觉过程。

DESCRIPTION (provided by applicant): The purpose of this project is to identify, that is, "clone," the RP10 gene which causes autosomal dominant retinitis pigmentosa (adRP). In preliminary research we localized the RP10 gene to human chromosome 7q31 by linkage mapping in 4 large, unrelated adRP families. The region containing the RP10 gene, flanked by markers D7S2471 and RP-STR12, is 3.3 mbp in length. By combining the public (GenBank) and private (Celera) human genome sequences, we have assembled 99% of the complete sequence in this region. Likewise, the Celera database contains 99% of the syntenic mouse sequence. By computational analysis and laboratory methods we have identified 38 independent, expressed sequences in the region, 10 of which are retinal-expressed and, thus, possible disease-gene candidates. We propose to complete the task of cloning the RP10 gene by building on these findings and resources. Proposed aims are 1) to reduce the RP10 linkage region, 2) to complete the physical and transcriptional maps of the region and 3) to test candidate genes in RP10 families. Methods for Aim 1 include enhanced linkage analysis and SNP haplotype testing to refine breakpoints; and production of haploid cell lines from patients to facilitate testing. Methods for Aim 2 include further computational analyses, comparing mouse and human genomes; and prioritization of candidate genes based on transcript profiles and function. Methods for Aim 3 include detection of retinal and photoreceptor-expressed genes in the region using knockout mice; candidate gene screening; and confirmation of positive results in unrelated adRP patients. These studies will be done in collaboration with the South African National Bioinformatics lnstitute, and the Ocular Genetics Unit, Trinity College, Dublin. Mutations in the RP10 gene may cause at least 10% of adRP which, in turn, affects thousands of Americans. Identification of the RP10 gene will provide direct benefits to affected individuals through diagnosis and counseling. Also, it is a necessary first step in designing gene-specific therapies and identifying modifying factors. More broadly, identification of genes causing retinitis pigmentosa has revealed new functional pathways in the retina, thus cloning the RP10 gene will likely contribute to a better understanding of normal visual processes.