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project/introduction.html

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-                                                &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;In order to achieve continuous directed evolution in nonconventional yeast strains, we try to construct the nyEvolvR system into K. marxianus. The nyEvolvR system integrates the specificity of the CRISPR/Cas9 system with the mutagenic activity of error-prone DNA polymerases. This approach utilizes a designed guide RNA (gRNA) of approximately 20 nucleotides for precise targeting. The nyEvolvR technology consists of two critical components: enhanced nick Cas9 (enCas9) and error-prone DNA polymerase I, which are fused together. The mechanism underlying the nyEvolvR system is straightforward: the gRNA guides enCas9 to the target motif, creating a gap. Subsequently, the error-prone DNA polymerase I fills the gap by introducing random base substitutions, leading to mutations in the target gene region and facilitating continuous directed evolution.
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+                                                &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;After constructing the cell factory, in order to obtain a higher yield strain, we tried to perform continuous directed evolution of the synthase of the target product. In a traditional directed evolution experiment, at least some of these steps are carried out manually, necessitating substantial investment of researcher time and lengthy evolution campaigns, which typically take months or longer to complete. Continuous evolution, by contrast, automates all of these steps, greatly reducing the generation time and the effort required from the researcher during evolution. These advantages enable large sequence-space searches, iterative improvements, and dramatic changes in phenotype over practical time scales. Continuous evolution also enables the exploration of longer and more numerous evolutionary trajectories, increasing the likelihood of accessing solutions that require many steps through sequence space and greatly facilitating the iterative refinement of selection conditions and targeted mutagenesis strategies.
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