Single Living Cell Analysis Nanoplatform for High-Throughput Interrogation of Gene Mutation and Cellular Behavior

Area of Science:

• Biotechnology
• Molecular Biology
• Cancer Research

Background:

• Genetic heterogeneity in cancer complicates precise drug treatment.
• Current single-cell gene analysis methods often destroy cells, limiting correlation with real-time behavior.
• Developing methods to link genetic profiles with cellular responses in living cells is crucial.

Purpose of the Study:

• To develop a high-throughput nanoplatform for analyzing gene properties and drug resistance in millions of single living cancer cells.
• To overcome the limitations of cell lysis in current single-cell analysis methods.
• To investigate the relationship between intracellular genetic properties and real-time cellular behaviors.

Main Methods:

• Designed a 'Domino-probe' for amplified fluorescence signal detection of intracellular target RNAs.
• Developed an on-chip addressable microwell-nanopore array for efficient probe delivery and cell observation.
• Utilized the nanoplatform for analyzing primary lung cancer cell samples.

Main Results:

• The nanoplatform successfully interrogated gene properties and drug resistance in millions of single cells.
• Demonstrated enhanced electro-delivery of the Domino-probe and in situ observation of cell behaviors.
• Validated a positive correlation between the ratio of EGFR mutant cells and their drug susceptibilities in lung cancer samples.

Conclusions:

• The developed nanoplatform enables high-throughput, precise analysis of single living cells.
• This technology facilitates the exploration of genotype-phenotype relationships at the single-cell level.
• The platform holds significant potential for advancing cancer research and personalized drug development.