Gene Knockout Cell Lines

Gene knockout stable cell lines are engineered cell lines where a specific gene has been permanently inactivated or "knocked out." This is typically achieved using gene-editing technologies such as CRISPR/Cas9 or TALENs. These cell lines provide a reliable and reproducible platform to study the complete loss-of-function effects of a gene, making them indispensable tools in functional genomics, drug discovery, and disease modeling.

Content

• Generation:
  • Target Selection: Designing gRNAs or TALENs specific to the gene of interest, minimizing off-target effects.
  • Gene Editing:
    • Transfection with plasmids or ribonucleoprotein (RNP) complexes containing Cas9 and gRNA.
• • Clonal Isolation: Single-cell cloning to isolate and expand knockout clones.
• Validation:
  • Genotypic Analysis:
    • PCR and sequencing of the target region to confirm indels or large deletions.
  • Phenotypic Analysis:
    • Western blot or immunofluorescence to verify the absence of the target protein.

Applications

• Functional Genomics:
  • Studying the role of genes in biological processes such as cell cycle regulation, differentiation, or apoptosis.
• Disease Modeling:
  • Mimicking loss-of-function mutations observed in genetic disorders, cancers, or metabolic diseases.
• Drug Discovery:
  • Validating therapeutic targets by assessing the impact of gene knockout on disease-relevant pathways.

Gene knockout stable cell lines are essential tools in modern molecular biology, offering robust and permanent platforms to investigate gene function, study disease mechanisms, and develop new therapeutic strategies.