The DOI matches its Crossref record; 71 of the 71 references it cites were found in a registry or an index.
Multiplexed and Programmable Regulation of Gene Networks with an Integrated RNA and CRISPR/Cas Toolkit in Human Cells
Open accessAbstract
Summary RNA-based regulation and CRISPR/Cas transcription factors (CRISPR-TFs) have yet to be integrated for multiplexed and tunable modulation of gene networks. Moreover, guide RNAs (gRNAs) for CRISPR-TFs have only been expressed from RNA polymerase III promoters in human cells, which limits their use for conditional gene regulation. To overcome these challenges, we combined multiple RNA regulatory strategies, including RNA-triple-helix structures, introns, microRNAs, and ribozymes, with Cas9-based CRISPR-TFs and Cas6/Csy4-based RNA processing in human cells. We describe three strategies for expressing functional gRNAs from RNA polymerase II promoters, which enable multiplexed production of proteins and multiple gRNAs from a single transcript. We used these tools to efficiently modulate endogenous promoters and implement tunable synthetic circuits, including multi-stage cascades and RNA-dependent networks that could be rewired with Csy4 to achieve complex behaviors. This multiplexable toolkit will be valuable for programming scalable gene circuits and perturbing endogenous networks for biology, therapeutic, and synthetic-biology applications.
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Cites
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Cited by
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Metadata
- Authors
- Nissim, L., Perli, S. D., Fridkin, A., Perez-Pinera, P., & Lu, T. K.
- Venue
- Molecular Cell
- Publisher
- Elsevier BV
- Published
- 2014-05
- Volume
- 54
- Issue
- 4
- Pages
- 698-710
- ISSN
- 1097-2765 (ISSN-L)
- OpenAlex
- W2028543816
- Cited by
- 438
- References
- 71
- FWCI
- 20.08
- Type
- journal article
- Language
- en
- Open access
- bronze
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- Nissim, L., Perli, S. D., Fridkin, A., Perez-Pinera, P., & Lu, T. K. (2014). Multiplexed and Programmable Regulation of Gene Networks with an Integrated RNA and CRISPR/Cas Toolkit in Human Cells. Molecular Cell. https://doi.org/10.1016/j.molcel.2014.04.022
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