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A Microfluidic Technique to Probe Cell Deformability
Published on: September 3, 2014
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Microfluidic Cell Deformability Assay for Rapid and Efficient Kinase Screening with the CRISPR-Cas9 System
Xin Han1,2, Zongbin Liu1,2, Li Zhao3
1Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX, 77030, USA.
Angewandte Chemie (International Ed. in English)
|June 4, 2016
Summary
This study used CRISPR-Cas9 gene editing and microfluidics to screen for kinases controlling cancer cell deformability and invasion. Loss of specific kinases increased cell flexibility and metastatic potential, identifying key regulators of tumor suppression.
Area of Science:
- Biotechnology
- Cancer Research
- Genetics
Background:
- Cancer cell deformability and invasiveness are critical for metastasis.
- Identifying genetic regulators of these mechanical properties is essential for understanding tumor progression.
Purpose of the Study:
- To perform a genome-wide CRISPR-Cas9 loss-of-function screen to identify kinases regulating cancer cell deformability and invasive potential.
- To develop and utilize a high-throughput microfluidic chip for cell mechanical property screening.
Main Methods:
- CRISPR-Cas9 screening was employed to generate kinase loss-of-function mutations in cancer cells.
- A microfluidic cell separation chip was used to isolate cells based on deformability.
- Deep sequencing was performed to identify genes associated with increased cell deformability and invasiveness.
Main Results:
- Loss of specific kinases significantly increased cancer cell deformability and invasive potential.
- Identified kinases include known tumor suppressors like CHK2, IKK-α, p38 MAPKs, and DAPK2.
- STK4 was validated as a key regulator of cell deformability and tumor suppression.
Conclusions:
- CRISPR-based on-chip mechanical screening is a powerful strategy for systematic genetic analysis of cell mechanics.
- This approach can identify novel regulators of cancer cell deformability and metastasis.
- The findings highlight the role of specific kinases in controlling cancer cell mechanical properties and tumor suppression.
