Quick Knockin Mouse Models (Neutral Locus Targeting: Rosa26, Hprt)

For academic research: Gene function studies by overexpression if Knockout has no or little phenotype Rescue experiment to validate specificity of the Knockout phenotype Protein structure and function studies Expression pattern analysis Expression of specific isoform to decipher its particular function Express human gene that has no mouse orthologue Model of dominant negative or gain-of-function mutation of human disease

For bio-pharmaceutical research & development: Validate drug candidate by expression of mutant form of receptor mimicking its role Validate mutation in target as disease-causing agent Test protective effect on controlled gene expression on disease onset or progression Identification biomarkers in human disease model Get mouse model that cross-reacts with human-specific therapeutic antibody (or similar)

+	Very robust approach due to the wide usage of both, Rosa26 and Hprt, by scientific community Fast development due to ready-to-use vectors Several genetic backgrounds are available (C57Bl/6, BALB/c…) Variety of genetic designs to fit a broad range of applications

-	No physiological regulation of the transgene expression →  Limitation can be bypassed by using conventional Knockin approach In most cases only one isoform can be expressed Mutated mouse gene is expressed in presence of the wildtype gene

Model recapitulating key features of human tauopathies.

Bondulich MK, et al. Tauopathy induced by low level expression of a human brain-derived tau fragment in mice is rescued by phenylbutyrate. Brain. 2016.

Tauopathies are characterized by progressive cognitive and/or motor dysfunction, together with highly phosphorylated aggregates of the microtubule-associated protein tau in brain and peripheral nerve.

Most existing mouse models of tauopathy overexpress mutant tau at levels that do not occur in human neurodegenerative disease.

Model: New highly disease-relevant mouse model of tauopathy expressing Hprt locus-targeted Tau35, highly phosphorylated C-terminal human tau fragment.

Aim: Evaluate a new model of tauopathy for developing novel treatments for human tauopathies.

Results: Tau35 mice represent a pathophysiologically relevant mouse model in which to test new, potentially disease-modifying therapies.

 

Figure 1. Tau expression in Tau35 mouse brains

Excessive accumulation of cGMP and subsequent rod photoreceptor death, followed by a mutation-independent, secondary death of cone photoreceptors.

A) RT-PCR confirms Tau35 expression.

B) Sagittal sections show widespread hemagglutinin labeling in Tau35 mouse brain (upper panels, scale bar = 2 mm). Higher magnifications of the hippocampal CA1 region show strongly hemagglutinin-positive pyramidal neurons in Tau35 mice (lower panels, scale bar = 200 mm). Western blots of frontal region and hippocampus/associated cortex (HC) show hemagglutinin protein expression only in Tau35 mice.

 

Figure 2. Progressive neuromuscular impairment

A) Limb clasping is apparent in Tau35 mice from 2 months of age (image shows 8 months), with all Tau35 animals affected by 18 months. Clasping is not observed in wildtype (wt) mice at any age examined.

B) Spine curvature is apparent in Tau35, but not wildtype mice, at 14 months of age. A progressive reduction in the kyphotic index in Tau35 mice after 4 months of age indicates increasing spine curvature.

C) Visible platform (VP) training in the Morris water maze was followed by 4 days of hidden platform training. At 10 months of age, Tau35 mice exhibit longer escape latency on the fourth day of testing compared to wildtype mice.

D) The grip strength of Tau35 mice declines steadily with age.

 

Figure 3. Phenylbutyrate rescues disease-related changes in Tau35 mice.

A) Morris water maze (10 months) testing of 4-phenylbutyrate (PBA, dotted lines) and vehicle-treated (solid lines) Tau35 (circles) and wildtype (wt, squares) mice. PBA-treated Tau35 mice show improved learning after 3 days, resulting in decreased escape latency.

B) Grip strength of Tau35 (10 months) is restored by PBA treatment.

Hprt | Rosa26

Adeline Sivignon, Mélissa Chervy, Caroline Chevarin, Elia Ragot, Elisabeth Billard, Jérémy Denizot, Nicolas Barnich.
An adherent-invasive Escherichia coli-colonized mouse model to evaluate microbiota-targeting strategies in Crohn's disease.
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Álvaro Sebastián-Serrano, Jesús Merchán-Rubira, Caterina Di Lauro, Carolina Bianchi, Lucía Soria-Tobar, Sonoko Narisawa, José L Millán, Jesús Ávila, Félix Hernández, Miguel Díaz-Hernández.
TNAP upregulation is a critical factor in Tauopathies and its blockade ameliorates neurotoxicity and increases life-expectancy
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Charlotte Douglas, Valdone Maciulyte, Jasmin Zohren, Daniel M Snell, Shantha K Mahadevaiah, Obah A Ojarikre, Peter J I Ellis, James M A Turner
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Faik Imeri, Bisera Stepanovska Tanturovska, Stephanie Schwalm, Sarbari Saha, Jinyang Zeng-Brouwers, Herrmann Pavenstädt, Josef Pfeilschifter, Liliana Schaefer, Andrea Huwiler.
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Lukas Huijbregts, Virginie Aiello, Andrea Soggia, Philippe Ravassard, Latif Rachdi, Raphaël Scharfmann, Olivier Albagli.
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Rosa26

Zhichao Ni, Lingxia Lyu, Hui Gong, Luping Du, Zuoshi Wen, Hua Jiang, Hao Yang, Yanhua Hu, Bohuan Zhang, Qingbo Xu, Xiaogang Guo, Ting Chen.
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Marc L Purazo, Ryan J Ice, Rahul Shimpi, Mark Hoenerhoff, Elena N Pugacheva.
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Modification of BRCA1-associated breast cancer risk by HMMR overexpression.
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Isabel Fernández-Pisonero, Laura Clavaín, Javier Robles-Valero, L Francisco Lorenzo-Martín, Rubén Caloto, Blanca Nieto, Carmen García-Macías, Clara L Oeste, Manuel Sánchez-Martín, Antonio Abad, Alejandro Hortal, Dolores Caballero, Marcos González, Mercedes Dosil, Balbino Alarcón, Xosé R Bustelo.
A hotspot mutation targeting the R-RAS2 GTPase acts as a potent oncogenic driver in a wide spectrum of tumors.
Cell Rep. 2022 Mar 15

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Valentina Poli, Marco Di Gioia, Martha Sola-Visner, Francesca Granucci, Andrew L Frelinger 3rd, Alan D Michelson, Ivan Zanoni.
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Yannick Fotio, Kwang-Mook Jung, Francesca Palese, Andre Obenaus, Alex Mabou Tagne, Lin Lin, Tarif Ibne Rashid, Romario Pacheco, Amandine Jullienne, Jade Ramirez, Marco Mor, Gilberto Spadoni, Cholsoon Jang, Andrea G Hohmann, Daniele Piomelli.
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