来自工程菌株的细胞自由菌体合成提高了产量
Rani Brooks1, Lisa Morici2, Nicholas Sandoval3
1Interdisciplinary Bioinnovation PhD Program, Tulane University, New Orleans, Louisiana 70118-5665, United States.
ACS synthetic biology
|August 7, 2023
概括
无细胞菌体合成 (CFBS) 通过使用工程细菌溶解物增强菌体T7的产生. 在转录/翻译机器 (TXTL) 中修改基因表达改善了高达10倍的菌体产量,加速了菌体治疗的发展.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 传染病研究 传染病研究
背景情况:
- 菌体疗法为抗药性感染提供了抗生素的有希望的替代方案,但面临着生产挑战.
- 无细胞菌体合成 (CFBS) 是一种体外生产菌体病毒的方法,克服了传统方法的局限性.
- 目前使用标准细菌溶解体的CFBS系统并没有针对高产量的菌体生产进行优化.
研究的目的:
- 通过无细胞菌体合成 (CFBS) 在体外增强菌体T7产量.
- 识别和操纵特定的细菌基因以优化转录/翻译机制 (TXTL) 以改善菌体的生产.
- 为了证明转基因细菌菌株在加速菌体制造方面的潜力.
主要方法:
- 利用可诱导的CRISPR干扰 (CRISPRi) 与核酶缺乏的Cas12a (dFnCas12a) 调节18个大肠杆菌BL21基因的表达.
- 过度表达的积极效应基因和抑制的负效应基因,以修改TXTL遗传背景.
- 通过CFBS使用优化的TXTL.产生的量化菌体T7产量.
主要成果:
- 工程化大肠杆菌BL21 TXTL显著增强了菌体T7 CFBS在体外的产量高达10倍.
- 过度表达翻译启动因子IF-3 (infC) 和小RNAOxyS和CyaR,提高了菌体产量.
- 抑制RecBCD外核酶的RecC子单元也导致了菌体生产的增加.
结论:
- 通过基因工程优化细菌TXTL是一种可行的策略,可以克服菌体制造瓶.
- 增强的CFBS具有显著的潜力,可以加速菌体治疗的发展和广泛采用.
- 这项研究为进一步改进无细胞菌体生产系统提供了基础.
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