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Multi-variable control to mitigate loads in CRISPRa networks: Extended Version

2024-09-11 · Krishna Manoj, Theodore W. Grunberg, Domitilla Del Vecchio

The discovery of CRISPR-mediated gene activation (CRISPRa) has transformed the way in which we perform genetic screening, bioproduction and therapeutics through its ability to scale and multiplex. However, the emergence of loads on the key molecular resources constituting CRISPRa by the orthogonal short RNA that guide such resources to gene targets, couple theoretically independent CRISPRa modules. This coupling negates the ability of CRISPRa systems to concurrently regulate multiple genes independent of one another. In this paper, we propose to reduce this coupling by mitigating the loads on the molecular resources that constitute CRISPRa. In particular, we design a multi-variable controller that makes the concentration of these molecular resources robust to variations in the level of the short RNA loads. This work serves as a foundation to design and implement CRISPRa controllers for practical applications.

📄 PDF Abstract BibTeX arXiv:2409.07384

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