Microfluidic Cell Deformability Assay for Rapid and Efficient Kinase Screening with the CRISPR-Cas9 System.
Herein we report a CRISPR-Cas9-mediated loss-of-function kinase screen for cancer cell deformability and invasive potential in a high-throughput microfluidic chip. In this microfluidic cell separation platform, flexible cells with high deformability and metastatic propensity flowed out, while stiff cells remained trapped. Through deep sequencing, we found that loss of certain kinases resulted in cells becoming more deformable and invasive. High-ranking candidates identified included well-reported tumor suppressor kinases, such as chk2, IKK-α, p38 MAPKs, and DAPK2. A high-ranking candidate STK4 was chosen for functional validation and identified to play an important role in the regulation of cell deformability and tumor suppression. Collectively, we have demonstrated that CRISPR-based on-chip mechanical screening is a potentially powerful strategy to facilitate systematic genetic analyses.
Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX, 77030, USA.|Department of Cell and Developmental Biology, Weill Medical College of Cornell University, New York, NY, 10065, USA.
Xin
X
Han
Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX, 77030, USA.|Department of Cell and Developmental Biology, Weill Medical College of Cornell University, New York, NY, 10065, USA.
Zongbin
Z
Liu
Department of Molecular and Human Genetics, Human Genome Sequencing Center, Baylor College of Medicine, Houston, TX, 77030, USA.
Li
L
Zhao
Department of Molecular and Human Genetics, Human Genome Sequencing Center, Baylor College of Medicine, Houston, TX, 77030, USA.
Feng
F
Wang
Department of Pediatrics, Baylor College of Medicine, Houston, TX, 77030, USA.
Yang
Y
Yu
Department of Pediatrics, Baylor College of Medicine, Houston, TX, 77030, USA.
Jianhua
J
Yang
Department of Molecular and Human Genetics, Human Genome Sequencing Center, Baylor College of Medicine, Houston, TX, 77030, USA.
Rui
R
Chen
Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX, 77030, USA. LQin@houstonmethodist.org.|Department of Cell and Developmental Biology, Weill Medical College of Cornell University, New York, NY, 10065, USA. LQin@houstonmethodist.org.
Lidong
L
Qin
D064113:CRISPR-Cas Systems / Q000235:genetics*; D045744:Cell Line, Tumor; D002465:Cell Movement; D064447:Checkpoint Kinase 2 / Q000378:metabolism; D015638:Cytochalasin D / Q000378:metabolism; D064550:Death-Associated Protein Kinases / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D056656:Lab-On-A-Chip Devices; D044085:Microfluidics; D008856:Microscopy, Fluorescence; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism; D000099317:Hippo Kinases
D064447:Checkpoint Kinase 2; D015638:Cytochalasin D; D064550:Death-Associated Protein Kinases; D004121:Dimethyl Sulfoxide; D051550:I-kappa B Kinase; D047908:Intracellular Signaling Peptides and Proteins; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C095566:STK4 protein, human; D000099317:Hippo Kinases
CRISPR-Cas9; analytical methods; biomarkers; cell deformability; microfluidics