Mesenchymal Stem Cells

Cas9 stable cell lines are engineered cell lines that stably express the Cas9 protein, derived from the CRISPR/Cas9 system. These cell lines are essential tools for genome editing applications, allowing researchers to introduce targeted genetic modifications such as knockouts, knock-ins, or transcriptional regulation.

Content on Cas9 Stable Cell Lines

• Generation:
  • Vector Design: Plasmids or viral vectors encoding the Cas9 gene are introduced into the host cells. Promoters like CMV or EF1α drive strong Cas9 expression.
  • Host Cells: Commonly used host cells include HEK293, CHO, and U2OS due to their compatibility with Cas9 and ease of culture.
  • Selection: Antibiotic resistance genes (e.g., puromycin, neomycin) or fluorescence-based markers (e.g., GFP) are used for selecting stable clones.
• Characteristics:
  • Stable Cas9 Expression: Enables consistent editing over multiple passages.
  • High Efficiency: Supports robust gene editing for various target loci.
  • Compatibility with gRNAs: Works seamlessly with transiently transfected or stably expressed guide RNAs (gRNAs).

Applications of Cas9 Stable Cell Lines

• Gene Knockout Studies:
  • Enable the generation of loss-of-function mutants by introducing gRNAs targeting specific genes.
  • Useful for studying gene function and pathway analysis.
• Gene Knock-In and Editing:
  • Facilitate the precise insertion or modification of genes using donor templates for homologous recombination (HR) or non-homologous end joining (NHEJ).
• CRISPR Interference (CRISPRi) and Activation (CRISPRa):
  • Modified Cas9 variants (dCas9) fused with transcriptional repressors or activators allow regulation of gene expression.

Cas9 stable cell lines are indispensable tools for CRISPR-based research and genome engineering, providing a robust and scalable platform for applications ranging from basic biology to therapeutic development.