Constitutive Knockout Cells | To get faster first results

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

Validation

Related resources and publications

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Reannotation of cancer mutations based on expressed RNA transcripts reveals functional non-coding mutations in melanoma

Am J Hum Genet

May 2025

Two birds with one stone: human SIRPα nanobodies for functional modulation and in vivo imaging of myeloid cells

Front Immunol

Dec 2023

Highlighted article

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CCX559 is a potent orally-administered small molecule PD-L1 inhibitor that induces anti-tumor immunity

PLoS One

Jan 2023

Highlighted article

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T cells expressing a chimeric-PD1-Dap10-CD3zeta receptor reduce tumor burden in multiple murine syngeneic models of solid cancer

Immunology

Jan 2020

ERα-36 regulates progesterone receptor activity in breast cancer

Breast Cancer Res

Jan 2020

Tumor Vessel Normalization, Immunostimulatory Reprogramming, and Improved Survival in Glioblastoma with Combined Inhibition of PD-1, Angiopoietin-2, and VEGF

Cancer Immunol Res

Jan 2019

SDHA gain-of-function engages inflammatory mitochondrial retrograde signaling via KEAP1-Nrf2.

Nat Immunol

Jan 2019

iPS technology: Four factors that will change the world… some day

iPS technology

5 min read

Nov 2017

Improving in vitro models: the coming of age of organoids

Organoids

6 min read

Mar 2018

A new promising family of small molecules regulating the PD-L1/PD-1 signaling pathway

Humanized PD-1/PD-L1 mouse and cell line

10 min read

Jul 2022

CCX559 is a potent, orally-administered small molecule PD-L1 inhibitor that induces anti-tumor immunity

Humanized ICP mouse and cell lines

9 min read

Jun 2023

Development of a new hSIRPα-specific radiotracer for non-invasive PET imaging using genOway’s double humanized genO-hCD47/hSIRPα mouse and genO-MC38-hPD-L1-hCD47-LZ cell line

Targeting the human CD47/SIRPα axis

10 min read

Mar 2024

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