유전자편집 기술을 활용한 난치 질환 유도 마우스 모델 제작 및 표현형 분석

연구책임자: 고려대학교 김경미 교수

세포와 세포소기관 유전자 염기교정 원천기술 개발과 고도화 연구

연구책임자: 울산대학교 김용섭

DddA-derived cytosine base editors (DdCBEs) and transcription activator-like effector (TALE)-linked deaminases (TALEDs) catalyze targeted base editing of mitochondrial DNA (mtDNA) in eukaryotic cells, a method useful for modeling of mitochondrial genetic disorders and developing novel therapeutic modalities. 

Here, we report that A-to-G-editing TALEDs but not C-to-T-editing DdCBEs induce tens of thousands of transcriptome-wide off-target edits in human cells. 

To avoid these unwanted RNA edits, we engineered the substrate-binding site in TadA8e, the deoxy-adenine deaminase in TALEDs, and created TALED variants with fine-tuned deaminase activity. 

Our engineered TALED variants not only reduced RNA off-target edits by >99% but also minimized off-target mtDNA mutations and bystander edits at a target site. 

Unlike wild-type versions, our TALED variants were not cytotoxic and did not cause developmental arrest of mouse embryos. 

As a result, we obtained mice with pathogenic mtDNA mutations, associated with Leigh syndrome, which showed reduced heart rates. 

Cell (2024)

https://www.sciencedirect.com/science/article/pii/S0092867423013211?via%3Dihub