In Vitro Generation of Genetic Diversity for Directed Evolution by Error-prone Artificial DNA Synthesis

Overview

Journal Commun Biol

Specialty Biology

Date 2024 May 24

PMID 38789612

Authors

Baowei Wang

Yang Liu

Xuelian Bai

Huijuan Tian

Lina Wang

Miao Feng

Hairong Xia

Affiliations

Soon will be listed here.

Abstract

Generating genetic diversity lies at the heart of directed evolution which has been widely used to engineer genetic parts and gene circuits in synthetic biology. With the ever-expanding application of directed evolution, different approaches of generating genetic diversity are required to enrich the traditional toolbox. Here we show in vitro generation of genetic diversity for directed evolution by error-prone artificial DNA synthesis (epADS). This approach comprises a three-step process which incorporates base errors randomly generated during chemical synthesis of oligonucleotides under specific conditions into the target DNA. Through this method, 200 ~ 4000 folds of diversification in fluorescent strength have been achieved in genes encoding fluorescent proteins. EpADS has also been successfully used to diversify regulatory genetic parts, synthetic gene circuits and even increase microbial tolerance to carbenicillin in a short time period. EpADS would be an alternative tool for directed evolution which may have useful applications in synthetic biology.

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