Related Experiment Video
Updated: Jun 17, 2026
10:13
A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells
Published on: July 3, 2013
11.6K
Inducible CRISPR-Cas9 screening platform to interrogate non-proliferative cellular states
Gabriele Casagrande Raffi1, Hendrik J Kuiken2, Cor Lieftink2
1Division of Molecular Carcinogenesis, Oncode Institute, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Nature Protocols
|October 8, 2025
Summary
This study introduces a new CRISPR screening protocol for non-proliferative cells. The inducible Cas9 system enables precise gene editing in states like senescence, aiding the discovery of new therapeutic targets.
Area of Science:
- Molecular Biology
- Genetics
- Cancer Research
Background:
- CRISPR screens are powerful tools for identifying genes essential for cell survival, proliferation, drug resistance, and synthetic lethal interactions.
- Applying loss-of-function CRISPR screening to non-proliferative cells is challenging due to slow editing and low sensitivity in detecting guide RNA dropout.
Purpose of the Study:
- To present a detailed protocol for performing CRISPR screens in non-proliferative states.
- To enable precise temporal control over gene editing using an inducible Cas9 system.
Main Methods:
- Development of an inducible Cas9 system for temporal control of gene editing.
- Utilizing flow cytometry to measure editing efficiency.
- Optimization of parameters for successful CRISPR screens in senescent cells.
Main Results:
- Demonstration of a robust protocol for CRISPR screening in non-proliferative cells.
- Successful application of the protocol to identify senolytic targets in senescent cells.
- The protocol is accessible and can be completed within 8-12 weeks.
Conclusions:
- The developed inducible CRISPR screening protocol overcomes limitations in studying non-proliferative cells.
- This method facilitates the identification of therapeutic targets in diverse fields, including aging and cancer.
- The protocol is adaptable for various non-proliferative states such as stem cell differentiation and immune cell development.
