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    CellREADR is a new technology that uses RNA sensing to control cell functions. This programmable system allows for easy monitoring and manipulation of animal cells, opening new avenues in biology and medicine.

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    Area of Science:

    • Molecular Biology
    • Biotechnology
    • Genetics

    Background:

    • Developing precise and scalable methods for accessing specific animal cell types is crucial for biological and medical research.
    • Existing technologies often lack the versatility and ease of implementation required for broad applications.

    Purpose of the Study:

    • To introduce and detail the CellREADR (Cell access through RNA sensing by Endogenous ADAR) technology.
    • To provide a comprehensive protocol for implementing CellREADR in both cell cultures and animal models.
    • To demonstrate the potential of RNA-based systems for cell monitoring and manipulation.

    Main Methods:

    • Designing and constructing CellREADR RNA sensor-actuator devices with specific RNA-binding regions and effector payloads.
    • Utilizing the ADAR (adenosine deaminase acting on RNA) editing mechanism for conditional translation of effector proteins.
    • Implementing protocols for validation in mammalian cell cultures and AAV-based delivery for in vivo studies, particularly in brain tissue.

    Main Results:

    • Successful design and validation of CellREADR in mammalian cell cultures within 2-3 weeks.
    • Demonstration of AAV-mediated in vivo delivery and characterization of CellREADR in animal models (6-8 weeks).
    • Established best practices, experimental controls, and troubleshooting for CellREADR implementation.

    Conclusions:

    • CellREADR offers a versatile, simple, and generalizable platform for RNA-based monitoring and manipulation of animal cells.
    • The technology leverages endogenous ADAR activity for programmable control, applicable across diverse tissues and species.
    • This protocol facilitates the broad application of CellREADR in fundamental research and therapeutic development.