TALE.Sense: A Versatile DNA Sensor Platform for Live Mammalian Cells.

Area of Science:

• Synthetic Biology
• Molecular Biology
• Genetics

Background:

• Developing precise tools for detecting specific DNA sequences in live cells is crucial for various biological applications.
• Existing DNA-sensing technologies, such as zinc-finger based sensors, have limitations in efficiency and dynamic range.

Purpose of the Study:

• To introduce TALE.Sense, a versatile platform for sensing DNA sequences in live mammalian cells.
• To demonstrate the programmable generation of custom cellular responses based on detected DNA targets.
• To compare the performance of TALE.Sense with existing DNA-sensing technologies.

Main Methods:

• Utilizing the programmable DNA-binding ability of transcription activator-like effectors (TALEs).
• Coupling TALE binding to conditional intein-reconstitution to create a trans-spliced ON-switch for a response circuit.
• Implementing transcriptional activation modules and SunTag-based amplification loops to enhance detection.

Main Results:

• TALE.Sense demonstrated higher efficiency and a broader dynamic range compared to zinc-finger based DNA sensors for the same DNA sequences.
• Modifications such as swapping transcriptional activation modules and adding SunTag amplification loops further improved detection efficiency.
• The platform exhibited versatility across a range of target sites.

Conclusions:

• TALE.Sense offers a versatile and efficient platform for sensing specific DNA sequences in live cells.
• Its ability to generate programmable responses makes it suitable for identifying live cell variants with desired DNA sequences.
• The platform can be integrated into synthetic biology circuits, expanding the possibilities for inducible genetic control.