Screening of small molecules targeting the splicing of SMN2 exon 7 to identify le

这个项目的目标是发现增加SMN2基因外显子7剪接的小分子。 前信使核糖核酸。全长SMN1转录本的产生可能会补偿SMN1的损失 脊髓性肌萎缩症患者的基因。之前对细胞和动物模型的研究表明， 增加人类SMN2基因剂量、转录速率和外显子7包涵体会导致SMN蛋白增加 并且可以补偿SMA中SMN1基因的丢失。通过高通量筛选 化合物文库我们已经鉴定了150个调节SMN2外显子7剪接的化合物，其中包括76个化合物 从组合化学库中衍生的化合物。我们现在建议探索结构类似物 在初步筛选中发现的这些化合物中，发现SMN2外显子更有效的调节子 7.拼接。这些化合物将成为开发SMA疗法的潜在先导化合物。 由于选择性前mRNA剪接的组合调节，靶向剪接的化合物 设备可能会影响多个外显子的剪接。这可能会导致不良的副作用和 排除某些化合物的治疗用途。为了解决这个问题，我们将确定全球影响 关于使用微阵列拼接每个化合物类。 我们的目标是： 目的1：通过筛选Smn2外显子7剪接的结构类似物来鉴定有效的调节子 在初步的高通量筛选实验中已经确定的化合物。放映将会是 使用高通量RT-PCR进行检测。 目标2：确定每一类代表性化合物的剪接的全局影响 微阵列。

The goal of this project is to discover small molecules that increase the splicing of exon 7 of the SMN2 gene pre-mRNA. The generation of full length SMN2 transcripts can potentially compensate the loss of the SMN1 gene in Spinal Muscular Atrophy (SMA) patients. Previous studies of cell and animal models show that increased human SMN2 gene dosage, transcription rate, and exon 7 inclusion result in elevated SMN protein levels and can compensate for the loss of the SMN1 gene in SMA. By high throughput screening of compound libraries we have identified 150 compounds modulating the splicing of SMN2 exon 7, including 76 compounds derived from combinatorial chemistry libraries. We now propose to explore structural analogues of these compounds identified in the preliminary screens, to discover more potent modulators of SMN2 exon 7 splicing. These will constitute potential lead compounds in the development of SMA therapeutics. Due to the combinatorial regulation of alternative pre-mRNA splicing, compounds targeting the splicing apparatus are likely to affect the splicing of a multiple exons. This may lead to undesired side effects and preclude the therapeutic use of certain compounds. To address this issue we will determine the global effect on splicing of each compound class using microarrays. Our aims are to: Aim 1: To identify potent modulators of SMN2 exon 7 splicing by screening structural analogues of compounds already identified in the preliminary high throughput screening experiments. The screening will be performed using high throughput RT-PCR. Aim 2: To determine the global effect on splicing of representative compounds from each class using microarrays.