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bioRxiv · 10.1101/2024.11.27.625622

De novo non-compartmentalized microorganism for directed evolution of nanobodies

Wu, D.·He, H.·Xi, R.·Xia, S.·Chen, X.
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Abstract

The creation of de novo microorganisms is a fascinating task, and microorganisms offer a wide range of applications for humanity. Herein, we designed a de novo microorganism, termed Syn-phage, as an artificial unencapsulated mimic of naturally occurring phage capable of evolving nanobodies with improved properties. Syn-phage composes unified genomic and phenotypic materials with phenotypic proteins encoded from its genetic information. Since Syn-phage can undergo amplification in E. coli. host, we established a protocol for the evolution of nanobodies with improved avidity or affinity, allowing super-resolution live-cell immunoimaging of endogenous proteins, a challenge posed in immunolabeling with antibodies. Utilizing a Syn-phage evolved nanobody, we visualized the microtubule nucleation factor hTPX2, recorded the dynamics of it during mitotic progression, and imaged it at a resolution beyond the diffraction limit of light via an AI-powered imaging pipeline. Hence, this study demonstrated the possibility to design de novo microorganisms for capitative applications.

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Wu, D., He, H., Xi, R., Xia, S., Chen, X.. 2024-11-28. De novo non-compartmentalized microorganism for directed evolution of nanobodies. https://doi.org/10.1101/2024.11.27.625622

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