category
NAR
date
Mar 24, 2026
slug
status
Published
summary
创新性地结合深度学习与湿实验多目标筛选流程,通过独立模型微调实现双适应性进化,显著提升T7 RNA聚合酶的热稳定性和高温活性(Tm提升>10°C,活性增强60倍),并降低副产物含量70%。
tags
蛋白质进化
核酸蛋白工具酶
type
Post

📄 原文题目

Deep learning-guided dual-fitness evolution of T7 RNA polymerase for enhanced stability and activity

🔗 原文链接

💡 AI 核心解读

创新性地结合深度学习与湿实验多目标筛选流程,通过独立模型微调实现双适应性进化,显著提升T7 RNA聚合酶的热稳定性和高温活性(Tm提升>10°C,活性增强60倍),并降低副产物含量70%。

📝 英文原版摘要

<span class="paragraphSection"><div class="boxTitle">Abstract</div>In protein engineering, simultaneously improving multiple fitness attributes is a critical yet challenging goal, largely due to the vastness of sequence space, the multifaceted interplay among different traits, and the complexity of non-linear mutational effects (epistasis). To address this, we developed a data-driven evolutionary strategy that couples <span style="font-style: italic;">in silico</span> deep learning with a wet-lab multi-objective selection workflow. By employing independent model fine-tuning for distinct traits, our approach facilitates navigating the fitness landscape to identify beneficial mutation combinations. We applied this strategy to T7 RNA polymerase (T7 RNAP), performing dual-fitness evolution to simultaneously enhance thermostability and activity at elevated temperatures. After five rounds of iterative evolution, we obtained T7 RNAP mutants exhibiting a melting temperature (<span style="font-style: italic;">T</span><sub>m</sub>) increase of &gt;10°C, a 60-fold enhancement in high-temperature activity, and a 70% reduction in by-product content. Validation in cell transfection demonstrated their potential for producing high-quality mRNA for industrial applications.</span>
利用DIAMOND DeepClust聚类生命蛋白宇宙阐明CRISPR–SaCas9动力学以实现有效HIV DNA切除的双gRNA策略
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