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.
Typical applications for constitutive Knockout cell lines
For academic research:
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For bio-pharmaceutical research & development:
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Strengths and limitations of constitutive Knockout cell lines
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- 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
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- 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
References
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.
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