Article
An evolved, orthogonal ssDNA generator for targeted hypermutation of multiple genomic loci.
Nucleic acids research - 24 Jan 2025
Chu Weiran, Tian Rongzhen, Guo Yaxin, Wu Yaokang, Rehm Fabian B H, Liu Long, Li Jianghua, Du Guocheng, Chen Jian, Liu Yanfeng
Abstract excerpt
Achieving targeted hypermutation of specific genomic sequences without affecting other regions remains a key challenge in continuous evolution. To address this, we evolved a T7 RNA polymerase (RNAP) mutant that synthesizes single-stranded DNA (ssDNA) instead of RNA in vivo, while still exclusively recognizing the T7 promoter. By increasing the error rate of the T7 RNAP mutant, it generates mutated ssDNA that...
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