Ann Hematol. 2019 Sep 9. doi: 10.1007/s00277-019-03763-2. [Epub ahead of print]

CRISPR/Cas9 gene correction of HbH-CS thalassemia-induced pluripotent stem cells.

Yingjun X¹, Yuhuan X¹, Yuchang C¹, Dongzhi L², Ding W¹, Bing S¹, Yi Y¹, Dian L¹, Yanting X¹, Zeyu X¹, Nengqing L¹, Diyu C¹, Xiaofang S³.

Abstract

Haemoglobin (Hb) H-constant spring (CS) alpha thalassaemia (- -/-α^CS) 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.