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Updated: May 10, 2026
Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
Published on: October 18, 2022
Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system.
David Bikard1, Wenyan Jiang, Poulami Samai
1Laboratory of Bacteriology, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA, Department of Microbiology and Immunobiology, Harvard Medical School, 4 Blackfan Circle, Boston, MA 02115, USA, Broad Institute of MIT and Harvard, 7 Cambridge Center, Cambridge, MA 02142, USA, McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA and Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Researchers engineered a Cas9 enzyme mutant to precisely control gene transcription. This technology enables programmable gene activation or repression, advancing synthetic biology and gene function studies.
Area of Science:
- Molecular Biology
- Synthetic Biology
- Gene Regulation
Background:
- Artificial control of transcription is crucial for understanding gene function and building synthetic gene networks.
- CRISPR-Cas9 is a bacterial immune system component, utilizing an RNA-guided DNA nuclease.
Purpose of the Study:
- To engineer a Cas9 nuclease mutant for programmable control of transcription.
- To develop methods for both gene activation and repression using modified Cas9.
Main Methods:
- Utilized a catalytically inactive Cas9 nuclease mutant with retained DNA-binding capabilities.
- Engineered Cas9 mutants to function as transcription repressors by blocking RNA polymerase binding.
- Developed a Cas9-omega subunit fusion for programmable transcription activation.
Main Results:
- Demonstrated Cas9 mutant's ability to act as a programmable transcription repressor.
- Successfully achieved programmable transcription activation through a Cas9 fusion protein.
- Showcased simple and efficient modulation of gene expression.
Conclusions:
- Engineered Cas9 mutants offer a versatile tool for precise gene expression control.
- This technology facilitates the study of gene networks and synthetic biology applications.
- Provides a foundation for developing novel biotechnological tools.
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