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Revision as of 20:50, 21 October 2019

2019 Team:Fudan-TSI

 

Construct your own mutation library

Mutation is the ever-powerful tool bringing diversity to the world and leading the way of evolution. Mutation library generation is critical for biological and medical research. Our system, R-Evolution, could easily establish the mutation library of a selected sequence, and allows for downstream high-throughput research applications such as directed evolution.

Take simple steps in vivo

Through the process of transcription, reverse transcription and recombination, the target sequence will be mutated in vivo.

Continuous evolution, consecutive selection

Iterative cycles of mutagenesis occur in one round of the system, allowing for high mutation rate. Selection couples with the system in one chassis.

Sequence specificity, host adaptability, high mutation rate and orthogonality

R-Evolution mutates only the target sequence, greatly reduces false-positive rate, uses orthogonal parts that functions independent of host’s environment, enables simultaneous scanning process in different hosts, goes through iterative rounds of mutagenesis with high efficiency, and couples mutagenesis with downstream operations.

Highlights of our project

project summary
Project by Team:Fudan-TSI

Mutation library generation is critical for biological and medical research, but current methods cannot mutate a specific sequence continuously without manual intervention. We hereby present a toolbox for in vivo continuous mutation library construction. First, the target DNA is transcribed into RNA. Next, our reverse transcriptase (RT) reverts RNA into cDNA, during which the target is randomly mutated by our RT's enhanced error-prone ability. Finally, the mutated version replaces the original sequence through recombination. These steps will be carried out iteratively, generating a random mutation library of the target with high efficiency as mutations accumulate along with bacterial growth. Our toolbox is orthogonal and provides a wide range of applications among various species. R-Evolution could mutate coding sequences and regulatory sequences, which enables the evolution of individual proteins or multiple targets at a time, promotes high-throughput research, and serves as a foundational advance to synthetic biology.