Point Mutation Cell Models | Focus on one function at a time

A point mutation Knockin cell line defines a model in which one or more nucleotides are constitutively mutated.

The insertion, deletion, nonsense and sense mutations alter the amino acid sequence of a given protein, thus changing its functions or properties.

Applications

For academic research:

Decipher protein function (gain or loss of function)‍
In vitro recapitulation of a human disease

For bio-pharmaceutical research & development:

Pharmacological off-target and efficacy studies by suppressing xenobiotics or antibody binding
Study drug-resistant mutants
Synthetic mortality screening in cancers

Strengths of point mutation Knockin cell models

High physiological relevancy of the scientific data obtained from the model (regulatory elements conserved, under control of endogenous promoter, expression of all splice variants, etc.) = more relevant than classical KO where the protein is absent
Reliable since each cell line is matched with an isogenic control cell clone

Limitations of point mutation Knockin cell models

Unpredicted alteration of splicing or 3D-protein structure

→ If the genetic modification creates a dominant mutation, an overexpression of the point mutant in a safe harbor site (Quick Knockin), might be a valuable option

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

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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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Point mutation KI cells

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Inducible Knockdown cells

Through a single genetic modification, the addition of a self-cleaving degron tag to a protein of interest allows its inducible, and reversible, degradation upon treatment.

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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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Quick KI cells

A quick Knockin cell line has a transgene inserted into permissive loci (e.g., Rosa26). The transgene overexpression is constitutively or inducibly.

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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.