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Multiple Input Sensing and Signal Integration Using a Split Cas12a System.
Molecular Cell ( IF 16.0 ) Pub Date : 2020-01-30 , DOI: 10.1016/j.molcel.2020.01.016 Hannah R Kempton 1 , Laine E Goudy 1 , Kasey S Love 1 , Lei S Qi 2
Molecular Cell ( IF 16.0 ) Pub Date : 2020-01-30 , DOI: 10.1016/j.molcel.2020.01.016 Hannah R Kempton 1 , Laine E Goudy 1 , Kasey S Love 1 , Lei S Qi 2
Affiliation
The ability to integrate biological signals and execute a functional response when appropriate is critical for sophisticated cell engineering using synthetic biology. Although the CRISPR-Cas system has been harnessed for synthetic manipulation of the genome, it has not been fully utilized for complex environmental signal sensing, integration, and actuation. Here, we develop a split dCas12a platform and show that it allows for the construction of multi-input, multi-output logic circuits in mammalian cells. The system is highly programmable and can generate expandable AND gates with two, three, and four inputs. It can also incorporate NOT logic by using anti-CRISPR proteins as an OFF switch. By coupling the split dCas12a design to multiple tumor-relevant promoters, we provide a proof of concept that the system can implement logic gating to specifically detect breast cancer cells and execute therapeutic immunomodulatory responses.
中文翻译:
使用拆分式Cas12a系统的多输入感测和信号集成。
适当时整合生物信号并执行功能响应的能力对于使用合成生物学进行复杂细胞工程至关重要。尽管CRISPR-Cas系统已被用于基因组的合成操作,但尚未完全用于复杂的环境信号传感,整合和激活。在这里,我们开发了一个分离的dCas12a平台,并表明它允许在哺乳动物细胞中构建多输入多输出逻辑电路。该系统具有高度可编程性,可以生成带有两个,三个和四个输入的可扩展AND门。通过使用抗CRISPR蛋白作为OFF开关,它也可以整合NOT逻辑。通过将分离的dCas12a设计与多个肿瘤相关启动子偶联,
更新日期:2020-02-06
中文翻译:
使用拆分式Cas12a系统的多输入感测和信号集成。
适当时整合生物信号并执行功能响应的能力对于使用合成生物学进行复杂细胞工程至关重要。尽管CRISPR-Cas系统已被用于基因组的合成操作,但尚未完全用于复杂的环境信号传感,整合和激活。在这里,我们开发了一个分离的dCas12a平台,并表明它允许在哺乳动物细胞中构建多输入多输出逻辑电路。该系统具有高度可编程性,可以生成带有两个,三个和四个输入的可扩展AND门。通过使用抗CRISPR蛋白作为OFF开关,它也可以整合NOT逻辑。通过将分离的dCas12a设计与多个肿瘤相关启动子偶联,