Gene Targeting of Eβ-iPSCs
Design of sgRNA and ssODN
ConstrucƟon and Cloning of sgRNA into Cas9 Expression Plasmid
Clonal Expansion and Verification
Genomic DNA
ssODN template
Annealed sgRNA oligo
sgRNA
Cas9
sgRNA/
PX459
sgRNA/
PX459
PX459
u6 promoter
Puromycin R
sgRNA
scaffold
SpCas9
u6 promoter
Puromycin R
sgRNA
scaffold
SpCas9
sgRNA
sgRNA
scaffold
sgRNA
SpCas9
u6 promoter
Puromycin R
Fig. 1 Schematic diagram of genome editing in Eβ-iPSCs
Correction of HbE/β-Thalassemia iPSCs using CRISPR/Cas9
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