Constitutive Knockout Cell Lines (Conventional)


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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.

Infographic: Conventional Knockout mouse model

Typical applications for constitutive Knockout cell lines

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 and limitations of constitutive Knockout cell lines

  • 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
  • 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



Henri-Philippe Konan, Loay Kassem, Soleilmane Omarjee, Ausra Surmieliova-Garnès, Julien Jacquemetton, Elodie Cascales, Amélie Rezza, Olivier Trédan, Isabelle Treilleux, Coralie Poulard, Muriel Le Romancer.
ERα-36 Regulates Progesterone Receptor Activity in Breast Cancer.
Breast Cancer Res. 2020 May 19.

Geoffrey Parriott, Kelsey Deal, Shane Crean, Elle Richardson, Emily Nylen, Amorette Barber.
T-cells Expressing a chimeric-PD1-Dap10-CD3zeta Receptor Reduce Tumour Burden in Multiple Murine Syngeneic Models of Solid Cancer.
Immunology. 2020 Mar 7.


Follow this link if you seek another customized cell line model.