한빛사 논문
Jeong Hyeon Lee,1,4 Hye-Kyung Oh,2,4 Beom Seok Choi,2 Ho Hyeon Lee,2 Kyu Jun Lee,2 Un Gi Kim,2 Jina Lee,2 Hyerim Lee,2 Geon Seong Lee,1 Se Jun Ahn,1 Jeong Pil Han,1 Seokjoong Kim,2 Su Cheong Yeom,1,3 and Dong Woo Song2
1Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, 1447 Pyeongchang-Ro, Daewha, Pyeongchang, Gangwon 25354, Korea;
2Research and Development Center, Toolgen Incorporated, Geumcheon-gu, Seoul 08501, Korea;
3WCU Biomodulation Major, Department of Agricultural Biotechnology, Seoul National University, Gwanank-gu, Seoul 08826, Korea
4These authors contributed equally
Correspondence: Su Cheong Yeom, PhD, Dong Woo Song, PhD
Abstract
Recently, clinical trials of adeno-associated virus-mediated replacement therapy have suggested long-term therapeutic effects for several genetic diseases of the liver, including hemophilia. However, there remain concerns regarding decreased therapeutic effects when the liver is regenerated or when physiological proliferation occurs. Although genome editing using the clustered regularly interspaced short palindromic repeats/Cas9 system provides an opportunity to solve this problem, low knock-in efficiency may limit its application for therapeutically relevant expression. Here, we identified a novel gene, APOC3, in which a strong promoter facilitated the expression of knocked-in genes in hepatocytes. We also investigated the effects of APOC3 editing using a small Cas9 protein derived from Campylobacter jejuni (CjCas9) in a hemophilic model. We demonstrated that adeno-associated virus-mediated delivery of CjCas9 and donor led to moderate levels of human factor 9 expression in APOC3-humanized mice. Moreover, knock-in-driven expression induced substantial recovery of clotting function in mice with hemophilia B. There was no evidence of off-target editing in vitro or in vivo. Collectively, our findings demonstrated therapeutically relevant expression using a precise and efficient APOC3-editing platform, providing insights into the development of further long-term therapeutics for diverse monogenic liver diseases.
논문정보
관련 링크
연구자 키워드
관련분야 연구자보기
관련분야 논문보기
해당논문 저자보기