Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510080, China.
Prenatal Diagnostic Centre, Guangzhou Women and Children Medical Centre affiliated to Guangzhou Medical University, Guangzhou, Guangdong, China.
Ann Hematol. 2019 Dec;98(12):2661-2671. doi: 10.1007/s00277-019-03763-2. Epub 2019 Sep 9.
Haemoglobin (Hb) H-constant spring (CS) alpha thalassaemia (- -/-α) is the most common type of nondeletional Hb H disease in southern China. The CRISPR/Cas9-based gene correction of patient-specific induced pluripotent stem cells (iPSCs) and cell transplantation now represent a therapeutic solution for this genetic disease. We designed primers for the target sites using CRISPR/Cas9 to specifically edit the HBA2 gene with an Hb-CS mutation. After applying a correction-specific PCR assay to purify the corrected clones followed by sequencing to confirm the mutation correction, we verified that the purified clones retained full pluripotency and exhibited a normal karyotype. This strategy may be promising in the future, although it is far from representing a solution for the treatment of HbH-CS thalassemia now.
血红蛋白 H-constant spring（CS）α地中海贫血（- -/-α）是中国南方最常见的非缺失型血红蛋白 H 疾病。基于 CRISPR/Cas9 的基因校正患者特异性诱导多能干细胞（iPSC）和细胞移植现在代表了这种遗传性疾病的治疗解决方案。我们使用 CRISPR/Cas9 为目标位点设计引物，以特异性编辑具有 Hb-CS 突变的 HBA2 基因。在应用校正特异性 PCR 分析纯化校正的克隆后，通过测序确认突变校正，我们验证了纯化的克隆保留了完全多能性并表现出正常的核型。尽管从现在开始，这种策略还远未代表治疗血红蛋白 H-CS 地中海贫血的解决方案，但它在未来可能具有广阔的应用前景。
