MOLECULAR GENETICS OF DELL7PLL.2 MICRODELETION SYNDROME

DELL7PLL.2 微缺失综合征的分子遗传学

• 批准号: 2201116
• 负责人: Pragna Patel
• 金额: $ 18.45万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-04-01 至 1996-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2201116
• 关键词: DNA; DNA footprinting; REM sleep; RNA splicing; SDS polyacrylamide gel electrophoresis; artificial chromosomes; autosomal dominant trait; biochemical evolution; blood; child behavior disorders; child mental disorders; child physical development; chromosome deletion; chromosome translocation; complementary DNA; congenital heart disorder; congenital oral /facial /cranial defect; cytogenetics; diagnosis design /evaluation; evaluation /testing; flow cytometry; gene deletion mutation; gene expression; gene rearrangement; genetic disorder; genetic library; genetic markers; genetic polymorphism; genome; human genetic material tag; human subject; hybrid cells; language development; mental retardation; messenger RNA; molecular cloning; molecular genetics; natural gene amplification; nervous system disorder; nucleic acid repetitive sequence; nucleic acid sequence; plasmids; polymerase chain reaction; psychosomatic disorders; pulsed field gel electrophoresis; restriction fragment length polymorphism; restriction mapping; southern blotting; speech disorder diagnosis; transfection /expression vector

Del(17)(p11.2) or Smith-Magenis syndrome (SMS) is a multiple congenital anomaly/mental retardation syndrome which is associated with an interstitial deletion of band p11.2 of the short arm of chromosome 17. The consistent clinical features of SMS patients include dysmorphic features, short stature, and developmental delay. Variable clinical features include cleft lip/palate, congenital heart defects, microcornea, sleep disturbances including absent REM sleep, signs of peripheral neuropathy, aggressive and self-destructive behavior. We have identified over 40 unrelated patients demonstrating del(17)(p11.2) by high resolution cytogenetics and established lymphoblastoid cell lines on these patients. DNA and cell lines are also available on one or both parents of the majority of these patients. The long term objective of this study is to understand the molecular basis of SMS. This application proposes to determine the approximate size of the deletion in key patients by dual beam laser flow cytometry. The parental origin and molecular extent of the deletions will be determined by Southern analysis with markers mapping to the deletion interval. A panel of somatic cell hybrids which include the affected chromosome 17 from key SMS deletion or translocation patients will be constructed. Overlapping yeast artificial chromosome clones and cosmids encompassing the deletion interval will be isolated and contigs established. Additional markers if essential will be created by Alu- and LINE-PCR of radiation hybrids retaining 17p11.2 sequences and by microdissection cloning of 17p11.2. A long range physical map of the region encompassing the deletion interval in the human genome will be constructed. The deletion breakpoints in SMS patients will be identified, cloned and the sequence determined. A genetic map of the syntonic regions in the mouse genome will be obtained using conserved sequences identified from these clones. Expressed sequences will be sought and appropriate cDNA libraries will be screened to identify candidate genes responsible for behavioral and REM sleep abnormalities, ocular, cardiac and craniofacial abnormalities and for neuropathy. The correlated physical and phenotype map of 17p11.2 and the identification of candidate genes will in the long term provide valuable information on the basis of these latter disorders and provide valuable reagents for the diagnosis, treatment and possibly correction of this syndrome.

Del(17)(p11.2) 或 Smith-Magenis 综合征 (SMS) 是一种多发性先天性疾病 与以下情况相关的异常/精神发育迟滞综合征 17号染色​​体短臂p11.2带间质缺失。 SMS 患者的一致临床特征包括畸形特征、 身材矮小，发育迟缓。 可变的临床特征包括 唇/腭裂、先天性心脏病、小角膜、睡眠障碍 包括缺乏快速眼动睡眠、周围神经病变的体征、攻击性和 自我毁灭的行为。 我们已识别出 40 多名无关患者 通过高分辨率细胞遗传学证明 del(17)(p11.2) 和 在这些患者身上建立了淋巴母细胞系。 DNA和细胞 其中大多数的父母之一或双方都可以使用线路 患者。 这项研究的长期目标是了解分子基础 短信。 该应用程序建议确定的大致尺寸 通过双束激光流式细胞术对关键患者进行缺失。 父母的 缺失的起源和分子范围将由 Southern 确定 使用映射到删除间隔的标记进行分析。 躯体面板 细胞杂交体，其中包括因关键 SMS 删除而受影响的 17 号染色体 或易位患者将被建造。 重叠酵母 包含缺失区间的人工染色体克隆和粘粒 将被分离并建立重叠群。 必要时附加标记 将通过保留 17p11.2 的辐射杂交体的 Alu- 和 LINE-PCR 产生 序列并通过显微切割克隆 17p11.2。 远距离物理 人类基因组中包含缺失区间的区域图 将被建造。 SMS 患者中的删除断点将是 鉴定、克隆并确定序列。 的遗传图谱 小鼠基因组中的同调区域将使用保守的方法获得 从这些克隆中鉴定出的序列。 将寻找表达的序列 并筛选合适的cDNA文库以鉴定候选基因 负责行为和快速眼动睡眠异常、眼部、心脏和 颅面异常和神经病变。 相关的物理和 17p11.2的表型图和候选基因的鉴定将在 从长远来看，在后者的基础上提供有价值的信息 为疾病的诊断、治疗和治疗提供有价值的试剂 可能会纠正这种综合症。