Rosa26 Targeted Gene Knockin | Quick overexpression solution

A Quick Knockin cell line defines a model in which a transgene is inserted in a permissive locus, also known as neutral locus or safe harbor, such as Rosa26. The transgene can be constitutively or inducibly overexpressed.

Targeting those loci enables full control of the copy number and avoids transgene silencing or deregulation of neighbouring genes.

Applications

For academic research:

Gene function studies
Study disease-causing transgene, mutated or not

For bio-pharmaceutical research & development:

Compound screening
Selectivity screening
Drug affinity testing
Compound profiling

Strengths of Quick Knockin cell models

Stable and predictable expression
Almost no position effect variegation
Almost no transgene silencing
No deregulation of neighbouring gene

Limitations of Quick Knockin cell models

Exogenous regulation may result in lower physiological relevancy
Limited physiological relevancy when multiple isoforms have been described
Problem of stoichiometry if interaction with other partners for signaling
→ Must be solved by an in-depth scientific analysis and a rationale risk assessment prior to model development, all included in our project feasibility assessment
Not functional for expression of non-dominant mutations
→ Choose alternative approaches such as, e.g., point mutation Knockin

Validation

Related resources and publications

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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

Apr 2023

Highlighted article

read the comment

ERα-36 regulates progesterone receptor activity in breast cancer

Breast Cancer Res

Aug 2020

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

Immunology

Mar 2020

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

Cancer Immunol Res

Oct 2019

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

Nat Immunol

Aug 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

No results

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Reporter KI cells

A reporter Knockin cell line is a useful model when there are no available antibodies for the target or when protein measurement is laborious and costly.

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Point mutation KI cells

A point mutation Knockin cell line has one or more mutated nucleotides that alter the protein amino acid sequence, changing its functions or properties.

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Constitutive KO cells

A constitutive Knockout cell line refers to a conventional KO; a model in which the target gene is permanently inactivated, and no protein is produced.