Cell engineering method using fluorogenic oligonucleotide signaling probes and flow cytometry.

Area of Science:

• Biotechnology and Genetic Engineering
• Cell Biology
• Molecular Biology

Background:

• Cell engineering and isolation are critical for biological research.
• Existing methods may have limitations in detecting and purifying cells based on specific gene expression.

Purpose of the Study:

• To introduce and describe Chromovert® Technology, a new cell engineering tool.
• To demonstrate its capability in detecting and purifying living cells based on gene expression.

Main Methods:

• Utilizes fluorogenic oligonucleotide signaling probes and flow cytometry.
• Detects and isolates individual living cells expressing transfected or endogenous genes.
• Employs novel plasmid-encoded sequences for mRNA tags and differentially-detectable fluorogenic probes for multiplexed detection.

Main Results:

• Successfully produced cell lines expressing diverse ion channel and membrane proteins (e.g., ENaC, NaV1.7, GABA_A receptors, CFTR, GPCRs).
• Demonstrated multiplexed detection and isolation of cells expressing multiple genes without gene-specific probes.
• Showcased the ability to engineer cells without reliance on leader sequences or chaperones.

Conclusions:

• Chromovert® Technology serves as a valuable research tool.
• Enables enrichment and isolation of cells engineered for specific gene expression.
• Offers a streamlined approach for cell engineering applications.