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

ABSTRACT: Haemoglobin (Hb) H-constant spring (CS) alpha thalassaemia (- -/-?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.

SUBMITTER: Yingjun X 

PROVIDER: S-EPMC6900276 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature