김윤정 (서울대학교 약학대학) | 한빛사논문 > 한빛사

한빛사논문

조회381

서울대학교 약학대학

CV File 125 kb

Nat. Commun., May 11 2024, 15 (1) | https://doi.org/10.1038/s41467-024-48111-9 Enhancing genome editing in hPSCs through dual inhibition of DNA damage response and repair pathways

Ju-Chan Park^1,2,6, Yun-Jeong Kim^1,2,6, Gue-Ho Hwang³, Chan Young Kang³, Sangsu Bae^3,4,5& Hyuk-Jin Cha¹

¹College of Pharmacy, Seoul National University, Seoul, Republic of Korea.

²College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Republic of Korea.

³Genomic Medicine Institute, Seoul National University College of Medicine, Seoul, Republic of Korea.

⁴Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Republic of Korea.

⁵Cancer Research Institute, Seoul National University College of Medicine, Seoul, Republic of Korea.

⁶These authors contributed equally: Ju-Chan Park, Yun-Jeong Kim.

Corresponding author

Correspondence to Hyuk-Jin Cha.

Abstract

Precise genome editing is crucial for establishing isogenic human disease models and ex vivo stem cell therapy from the patient-derived hPSCs. Unlike Cas9-mediated knock-in, cytosine base editor and prime editor achieve the desirable gene correction without inducing DNA double strand breaks. However, hPSCs possess highly active DNA repair pathways and are particularly susceptible to p53-dependent cell death. These unique characteristics impede the efficiency of gene editing in hPSCs. Here, we demonstrate that dual inhibition of p53-mediated cell death and distinct activation of the DNA damage repair system upon DNA damage by cytosine base editor or prime editor additively enhanced editing efficiency in hPSCs. The BE4stem system comprised of p53DD, a dominant negative p53, and three UNG inhibitor, engineered to specifically diminish base excision repair, improves cytosine base editor efficiency in hPSCs. Addition of dominant negative MLH1 to inhibit mismatch repair activity and p53DD in the conventional prime editor system also significantly enhances prime editor efficiency in hPSCs. Thus, combined inhibition of the distinct cellular cascades engaged in hPSCs upon gene editing could significantly enhance precise genome editing in these cells.

논문정보

형식Research article
게재일2024년 05월 (BRIC 등록일 2024-05-13)
연구진국내 연구진
분야 의약학 > 재생의학

댓글 작성 로그인

CV 열람하기

연락처 보기