Enhanced homology-directed repair for highly efficient gene editing in hematopoietic stem/progenitor cells.
Suk See De RavinJulie BraultRonald J MeisSiyuan LiuLinhong LiMara Pavel-DinuCicera R LazzarottoTaylor Q LiuSherry M KoontzUimook ChoiColin L SweeneyNarda TheobaldGaHyun LeeAaron B ClarkSandra Sczerba BurkettBenjamin P KleinstiverMatthew H PorteusShengdar TsaiDouglas B KuhnsGary A DahlStephen J HeadeyXiaolin WuHarry L MalechPublished in: Blood (2021)
Lentivector gene therapy for X-linked chronic granulomatous disease (X-CGD) has proven to be a viable approach, but random vector integration and subnormal protein production from exogenous promoters in transduced cells remain concerning for long-term safety and efficacy. A previous genome editing-based approach using Streptococcus pyogenes Cas9 mRNA and an oligodeoxynucleotide donor to repair genetic mutations showed the capability to restore physiological protein expression but lacked sufficient efficiency in quiescent CD34+ hematopoietic cells for clinical translation. Here, we report that transient inhibition of p53-binding protein 1 (53BP1) significantly increased (2.3-fold) long-term homology-directed repair to achieve highly efficient (80% gp91phox+ cells compared with healthy donor control subjects) long-term correction of X-CGD CD34+ cells.
Keyphrases
- highly efficient
- induced apoptosis
- genome editing
- crispr cas
- cell cycle arrest
- endoplasmic reticulum stress
- gene expression
- signaling pathway
- oxidative stress
- rheumatoid arthritis
- staphylococcus aureus
- transcription factor
- escherichia coli
- pseudomonas aeruginosa
- small molecule
- candida albicans
- cell proliferation
- biofilm formation
- nk cells
- interstitial lung disease