DETECTION OF SINGLE BASE MUTATIONS IN RNA MOLECULES

RNA 分子中单碱基突变的检测

• 批准号: 3462545
• 负责人: FRANCES SMITH
• 金额: $ 9.58万
• 依托单位: MOUNT SINAI SCHOOL OF MEDICINE OF CUNY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-04-01 至 1992-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3462545
• 关键词: Gaucher's disease; gel electrophoresis; genetic disorder diagnosis; genetic mapping; messenger RNA; molecular genetics; nucleic acid hybridization; nucleic acid probes; nucleic acid sequence; oligonucleotides; phenylketonurias; point mutation

There is no rapid screening method available which reliably detects all point mutations in RNA molecules. The first part of this project is designed to improve current technologies in order to achieve this goal. Specifically, experiments will be designed to analyze RNA-RNA heteroduplexes in denaturing gradient gel electrophoresis and in solution in order to allow the rapid detection of mutations in mRNA molecules. In the second part of this grant, these techniques will be used to identify the mutations associated with Gaucher disease and phenylketonuria. The mutations will then be further characterized by directly sequencing the area of the RNA to which they have been localized. Subsequently, oligonucleotide probes will be synthesized and used to rapidly screen RNAs obtained from a large number of affected individuals, thus determining the frequency of each sequenced mutation in the affected population. Results obtained in these studies should contribute greatly to a better understanding of the genetic heterogeneity of genetic diseases and should also help us to correlate mutations with specific clinical disease patterns.

目前还没有可靠的快速筛查方法 RNA分子中的所有点突变。 第一部分 该项目旨在改进现有技术， 实现这一目标。 具体而言，实验将被设计为 在变性梯度凝胶中分析RNA-RNA异源双链体 电泳和溶液中，以允许快速 检测mRNA分子中的突变。 第二部分 这项授权，这些技术将被用来识别突变， 与戈谢病和苯丙酮尿症有关。 突变 然后将通过直接测序该区域来进一步表征 它们所定位的RNA。 随后，委员会注意到， 寡核苷酸探针将被合成并用于快速地 筛选从大量受影响个体获得的RNA， 从而确定每个测序的突变的频率， 受影响的人口。 这些研究的结果应 有助于更好地理解遗传学 遗传疾病的异质性，也应该帮助我们 将突变与特定的临床疾病模式相关联。