한빛사 논문
Hyeon-Ki Jang1,†,‡, Dong Hyun Jo2,†, Seu-Na Lee3,†, Chang Sik Cho4, You Kyeong Jeong1, Youngri Jung1, Jihyeon Yu1,§, Jeong Hun Kim4,5,6,*, Jae-Sung Woo3,*, Sangsu Bae1,*
1Department of Chemistry and Research Institute for Convergence of Basic Sciences, Hanyang University, Seoul 04763, South Korea. 2Department of Anatomy and Cell Biology, Seoul National University College of Medicine, Seoul 03080, South Korea. 3Department of Life Sciences, Korea University, Seoul 02841, South Korea. 4Fight against Angiogenesis-Related Blindness (FARB) Laboratory, Biomedical Research Institute, Seoul National University Hospital, Seoul 03080, South Korea. 5Department of Ophthalmology, Seoul National University College of Medicine, Seoul 03080, South Korea. 6Advanced Biomedical Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), Daejeon 34141, South Korea.
*Corresponding author.
†These authors contributed equally to this work.
‡Present address: Stem Cell Convergence Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), Daejeon 34141, South Korea.
§Present address: Division of Life Science, Korea Polar Research Institute, Incheon 21990, South Korea
Abstract
Ribonucleoprotein (RNP) complex–mediated base editing is expected to be greatly beneficial because of its reduced off-target effects compared to plasmid- or viral vector–mediated gene editing, especially in therapeutic applications. However, production of recombinant cytosine base editors (CBEs) or adenine base editors (ABEs) with ample yield and high purity in bacterial systems is challenging. Here, we obtained highly purified CBE/ABE proteins from a human cell expression system and showed that CBE/ABE RNPs exhibited different editing patterns (i.e., less conversion ratio of multiple bases to single base) compared to plasmid-encoded CBE/ABE, mainly because of the limited life span of RNPs in cells. Furthermore, we found that off-target effects in both DNA and RNA were greatly reduced for ABE RNPs compared to plasmid-encoded ABE. We ultimately applied NG PAM–targetable ABE RNPs to in vivo gene correction in retinal degeneration 12 (rd12) model mice.
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