A constitutive Knockout cell line, also referred to as conventional Knockout (KO), defines a model in which the target gene is permanently inactivated.

No protein is produced.

Applications

For academic research:

  • Determine and study main functions of the gene and/or protein
  • Loss-of-gene-function studies
  • In vitro recapitulation of a human disease

For bio-pharmaceutical research & development:

  • In vitro target validation
  • Mimic human disease for drug studies and screenings
  • Positive control for antagonist drug screening
  • Synthetic lethality screening
  • Study safety, specificity, and off-target activity of drug candidates
  • Improve performance of bioproduction models

Strengths of constitutive Knockout cell models

  • Total absence of protein including all isoforms, or total absence of a specific isoform
  • Reliable: each cell line is matched with an isogenic control cell clone
  • Feasible in all genetic backgrounds and cell types

Limitations of constitutive Knockout cell models

  • Functions of the KO protein may be compensated by related proteins, called genetic compensation
  • Genetic modification of the gene of interest may deregulate neighboring genes, inducing a non-specific phenotype
  • Challenging in cells with high polyploidy

→  Risks can be strongly minimized by applying in-depth bio-informatic, genetic, and bibliographic analyses
→ An alternative model is a functional Knockout by introducing a
point mutation to produce an inactive protein

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