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通过Cas9介导的局部和突变发生的BCL11A增强剂剖析

Matthew C Canver¹, Elenoe C Smith¹, Falak Sher¹

¹Division of Hematology/Oncology, Boston Children's Hospital, Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Stem Cell Institute, Department of Pediatrics, Harvard Medical School, Boston, Massachusetts 02115, USA.

|September 17, 2015

概括

研究人员使用集成的CRISPR- Cas9查绘制了BCL11A红细胞增强剂的关键特征. 这项研究验证了增强剂作为重新诱导胎儿血红蛋白的目标,为未来的基因组编辑疗法提供了信息.

科学领域:

基因组学
分子生物学
发育生物学

背景情况:

增强剂对于细胞身份至关重要,通常通过染色体标记和功能增益研究来识别.
功能丧失研究对于在基因组背景下确认增强剂的要求至关重要.
此前已经确定了与胎儿血红蛋白水平相关的BCL11A红色素增强剂,其对红色素BCL11A表达至关重要.

研究的目的:

使用聚合的CRISPR- Cas9指导RNA库进行人体和小鼠BCL11A红细胞增强剂的局部和突变发生.
确定这些增强剂的关键最小特征和漏洞.
验证BCL11A红细胞增强剂作为胎儿血红蛋白再诱导的治疗标.

主要方法:

开发和应用聚类定期间隔的帕林德罗姆重复 (CRISPR) -Cas9指导RNA库用于和突变发生.
人类和小鼠增强剂的现场功能审讯.
编辑人类原始基因和小鼠转基因进行验证.

主要成果:

确定BCL11A红色素增强剂中的关键最小序列和离散漏洞.
人类和小鼠增强剂之间保留功能的示范,但结构不同.
通过实验编辑和转基因来验证BCL11A红细胞增强剂作为胎儿血红蛋白再诱导的标.

结论:

这种BCL11A红细胞增强剂具有关键特征,其中一些特征似乎是灵长类的特征.
红细胞增强剂BCL11A是治疗基因组编辑的有效标,旨在重新诱导胎儿血红蛋白.
开发的选方法广泛适用于非编码基因组元素的功能分析.

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