The DOI matches its Crossref record; 61 of the 61 references it cites were found in a registry or an index.
Tunable and Multifunctional Eukaryotic Transcription Factors Based on CRISPR/Cas
Open accessAbstract
Transcriptional regulation is central to the complex behavior of natural biological systems and synthetic gene circuits. Platforms for the scalable, tunable, and simple modulation of transcription would enable new abilities to study natural systems and implement artificial capabilities in living cells. Previous approaches to synthetic transcriptional regulation have relied on engineering DNA-binding proteins, which necessitate multistep processes for construction and optimization of function. Here, we show that the CRISPR/Cas system of Streptococcus pyogenes can be programmed to direct both activation and repression to natural and artificial eukaryotic promoters through the simple engineering of guide RNAs with base-pairing complementarity to target DNA sites. We demonstrate that the activity of CRISPR-based transcription factors (crisprTFs) can be tuned by directing multiple crisprTFs to different positions in natural promoters and by arraying multiple crisprTF-binding sites in the context of synthetic promoters in yeast and human cells. Furthermore, externally controllable regulatory modules can be engineered by layering gRNAs with small molecule-responsive proteins. Additionally, single nucleotide substitutions within promoters are sufficient to render them orthogonal with respect to the same gRNA-guided crisprTF. We envision that CRISPR-based eukaryotic gene regulation will enable the facile construction of scalable synthetic gene circuits and open up new approaches for mapping natural gene networks and their effects on complex cellular phenotypes.
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Cited by
The next generation of CRISPR–Cas technologies and applications
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Beyond editing: repurposing CRISPR-Cas9 for precision genome regulation and interrogation
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The CRISPR-Cas immune system: Biology, mechanisms and applications
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Editing the epigenome: technologies for programmable transcription and epigenetic modulation
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Metadata
- Authors
- Farzadfard, F., Perli, S. D., & Lu, T. K.
- Venue
- ACS Synthetic Biology
- Publisher
- American Chemical Society (ACS)
- Published
- 2013-09-11
- Volume
- 2
- Issue
- 10
- Pages
- 604-613
- ISSN
- 2161-5063 (ISSN-L)
- OpenAlex
- W1968165219
- Cited by
- 358
- References
- 61
- FWCI
- 15.33
- Type
- journal article
- Language
- en
- Open access
- hybrid
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