Continuous genetic recording with self-targeting CRISPR-Cas in human cells

Area of Science:

• Molecular Biology
• Synthetic Biology
• Biotechnology

Background:

• Monitoring in vivo molecular events is crucial for understanding cellular signaling and behavior.
• Existing methods lack the capacity for longitudinal recording within cellular niches.

Purpose of the Study:

• To develop a self-contained analog memory device for in vivo recording of molecular stimuli into DNA mutations.
• To create a tool for longitudinal monitoring of cellular behavior and signaling dynamics.

Main Methods:

• A novel device using self-targeting guide RNA (stgRNA) and Streptococcus pyogenes Cas9 nuclease was engineered.
• The system enables localized, continuous DNA mutagenesis based on stgRNA expression.
• Programmable and multiplexed memory storage was demonstrated in human cells, triggered by inducers or inflammation.

Main Results:

• The Mammalian Synthetic Cellular Recorder Integrating Biological Events (mSCRIBE) was successfully developed and validated.
• Demonstrated in vitro and in vivo functionality in human cells.
• Showcased programmable and multiplexed data storage capabilities.

Conclusions:

• mSCRIBE offers a novel strategy for investigating cell biology in vivo.
• The device enables continuous evolution of targeted DNA sequences for biological recording.
• This technology advances the field of in vivo cellular monitoring and synthetic biology.