Cystic Fibrosis Animal Model Service
Cystic fibrosis (CF) is a life-threatening monogenic disorder driven by mutations in the CFTR gene, presenting significant challenges in translational drug development. At Protheragen, we provide custom cystic fibrosis animal model Services tailored to your specific research objectives. From targeted gene editing strategies to comprehensive phenotyping, we develop and characterize CF animal models that faithfully recapitulate the hallmarks of the disease, enabling rigorous preclinical evaluation of novel therapeutics. Whether you require mutation-matched models for modulator testing, conditional systems for organ-specific CFTR studies, or species-appropriate platforms for pulmonary and gastrointestinal endpoint assessment, our custom capabilities deliver the precision and scalability you need.
Overview of Cystic Fibrosis Animal Models
Cystic fibrosis is a recessive genetic disorder caused by pathogenic mutations in the CFTR gene, which encodes a chloride and bicarbonate ion channel critical for epithelial fluid and electrolyte homeostasis. Impaired CFTR function results in abnormal ion transport across epithelial surfaces, leading to delayed mucus clearance, chronic infection, progressive lung function decline, pancreatic insufficiency, gastrointestinal obstruction, and reproductive tract abnormalities.
Fig.1 Generation, expression, and function of the W1282X CFTR mouse model. (Michicich, M., et al., 2025)
The complex, multi-organ nature of CF necessitates the use of animal models that can faithfully recapitulate disease pathogenesis at both the molecular and physiological levels. Animal models of CF, particularly those generated through targeted gene editing, have become indispensable tools for investigating CFTR biology, elucidating mutation-specific disease mechanisms, and screening novel therapeutic candidates including CFTR modulators, readthrough agents, and gene-editing platforms.
Our Services
With deep expertise in genetic engineering and a rigorous focus on translational relevance, Protheragen offers a comprehensive suite of CF animal model services spanning the full preclinical development continuum. Our advantages include proficiency in generating mutation-specific models carrying clinically relevant CFTR alleles, robust phenotyping platforms integrating electrophysiological, histological, and biochemical assessments, and flexible service modules that accommodate custom model generation, in-life efficacy studies, and longitudinal disease monitoring. From establishing foundation breeding colonies to executing IND-enabling studies, our integrated approach ensures seamless translation from model characterization to investigational new drug application support.
Animal Models of Cystic Fibrosis
Protheragen provides custom development services for a wide range of CF animal models tailored to your specific mutation of interest, species preference, and experimental endpoints. Rather than maintaining a fixed inventory, we engineer and characterize models on a project basis using state-of-the-art gene editing technologies. The following categories and specific model types are available for custom generation, each selected for their established relevance in CF research.
Custom-generated models carrying precise CFTR mutations, including knockouts, patient-relevant point mutations, and conditional alleles. These models are fully characterized for CFTR expression, ion transport deficits, and multi-organ pathology.
Optional models:
- CFTR Knockout Model
- CFTR-G542X Mouse Model
- CFTR-W1282X Mouse Model
- CFTR-F508del Mouse Model
- CFTR-G551D Mouse Model
- Others
Mouse Model for Cystic Fibrosis Research
| Model Name | Modeling Method | Detailed Description |
|---|---|---|
| CFTR-KO Mouse Model | Knockout (KO) | The CFTR-KO mouse model was generated using gene-editing technology to achieve a targeted deletion of exons 5 and 6 within the CFTR gene. |
| CFTR-Flox | CKO | A conditional knockout model in which loxP sites were inserted flanking critical exons of the CFTR gene, enabling tissue-specific CFTR ablation upon Cre recombinase expression. |
| CFTR-G542X Mouse Model | Knock-in | A knock-in model carrying the patient-relevant G542X nonsense mutation in the endogenous CFTR locus, resulting in a premature termination codon and severely reduced CFTR protein expression. |
| CFTR-F508del Mouse Model | Knock-in | A knock-in model harboring the common F508del mutation, which leads to defective CFTR processing, trafficking, and channel gating. |
| CFTR-W1282X Mouse Model | Knock-in | A knock-in model carrying the W1282X nonsense mutation, engineered to recapitulate severe intestinal and respiratory phenotypes. |
| CFTR-G551D Mouse Model | Knock-in | A knock-in model with the G551D gating mutation, which impairs CFTR channel opening while preserving normal protein synthesis and trafficking, useful for potentiator compound evaluation. |
| … | … | … |
Case Study-CFTR-KO Mouse Model Development
Model Introduction
The CFTR-KO mouse model was developed as a complete loss-of-function platform for cystic fibrosis research. By disrupting the murine Cftr gene at exons 5 and 6, this model eliminates functional CFTR protein expression. Homozygous CFTR-KO mice recapitulate key CF manifestations including intestinal obstruction, mucus accumulation in the gastrointestinal and respiratory tracts, and reduced postnatal survival. This model serves as a foundational tool for studying CF disease mechanisms, evaluating gene replacement strategies, and testing mutation-agnostic therapeutic interventions.
Methodology
- Animal Model: A CFTR-KO mouse strain was generated using gene-editing technology to delete exons 5 and 6 of the endogenous Cftr gene. The targeting strategy was designed to remove critical coding regions, thereby disrupting the reading frame and preventing full-length CFTR protein synthesis. Heterozygous and homozygous knockout animals were produced through intercrossing, and genotyping was performed to confirm the deletion. All mice were maintained under controlled environmental conditions. For the survival study, a subset of homozygous CFTR-KO pups received prophylactic PEG treatment to alleviate intestinal obstruction.
- Phenotypic Analysis Methods: Tissue samples (large intestine, small intestine, lung, and trachea) were collected from 3-week-old homozygous CFTR-KO mice, heterozygous littermates, and wild-type controls (n=5 per group). Total RNA was extracted and reverse-transcribed, and quantitative RT-qPCR was performed to measure murine Cftr (mCFTR) transcript levels using gene-specific primers. For histological evaluation, intestinal, tracheal, and lung tissues were fixed, embedded in paraffin, sectioned, and stained with Alcian Blue-Periodic Acid Schiff (AB-PAS) to visualize neutral and acidic mucosubstances. Survival was monitored daily from birth, and survival curves were generated for untreated and PEG-treated homozygous CFTR-KO groups.
Phenotypic Analysis & Results
Gene expression analysis confirmed the absence of Cftr transcripts in homozygous CFTR-KO mice. RT-qPCR was performed on large intestine, small intestine, and lung tissues from 3-week-old mice. In wild-type and heterozygous CFTR-KO mice, mCFTR transcripts were readily detected in all three tissues. In contrast, no mCFTR transcripts were detected in any of the examined tissues from homozygous CFTR-KO mice, confirming complete loss of Cftr gene expression.
Fig.2 Gene expression detection in the large intestine, small intestine, and lung tissues of homozygous CFTR-KO mice and wild-type (WT) mice. Data are presented as mean ± SEM (n=5).
Homozygous CFTR-KO mice exhibited reduced survival and pronounced mucus pathology in both the intestinal and respiratory tracts. Survival monitoring revealed that homozygous CFTR-KO mice began to die as early as 2 weeks of age, and prophylactic PEG treatment significantly improved survival rates. AB-PAS staining of small intestine sections from 3-week-old homozygous CFTR-KO mice showed extensive mucus accumulation within intestinal glands, enlarged goblet cells, increased luminal and inter-villus mucus, and dilated intestinal glands compared to wild-type controls, which displayed only minimal mucus in the intestinal lumen and between villi. In the trachea, homozygous CFTR-KO mice exhibited dilated submucosal glands and increased intraglandular mucus substances. In the lung, while wild-type mice rarely showed mucus in the bronchial lumen, most homozygous CFTR-KO lung samples demonstrated marked mucus accumulation within the bronchial lumen, indicative of defective airway mucus clearance.
Fig.3 Survival analysis of homozygous CFTR-KO mice with or without prophylactic PEG treatment. Data are presented as mean ± SEM (n=10).
Conclusion
The CFTR-KO mouse model was successfully generated with a targeted deletion of exons 5-6, resulting in complete loss of Cftr transcript expression in all examined tissues. Homozygous knockout mice recapitulated hallmark CF pathologies including reduced survival, severe intestinal mucus accumulation with glandular dilation and goblet cell hypertrophy, and abnormal mucus phenotypes in the trachea and lung. This model provides a robust platform for mechanistic studies of CF pathophysiology and for preclinical evaluation of gene replacement, readthrough, and other mutation-agnostic therapeutic modalities.
Contact Us
Protheragen offers integrated preclinical support including pharmacokinetic studies, pharmacodynamic assessments, and safety pharmacology evaluations. Our portfolio encompasses in-life efficacy studies using clinically relevant endpoints such as nasal potential difference measurements, short-circuit current analyses, and organoid-based functional assays; longitudinal disease monitoring with multi-parameter phenotyping; and customized study designs tailored to candidate therapeutics including CFTR modulators, gene therapies, readthrough agents, and RNA-based platforms. Backed by rigorous quality standards and a science-driven approach, we partner with you to accelerate your CF therapeutic pipeline from discovery through regulatory submission. For a detailed discussion of your specific project requirements or to request a customized service proposal, please contact us.
Reference
- Michicich, Margaret et al. "A W1282X cystic fibrosis mouse allows the study of pharmacological and gene-editing therapeutics to restore CFTR function." Journal of cystic fibrosis: official journal of the European Cystic Fibrosis Society 24.1 (2025): 164-174.
All of our services and products are intended for preclinical research use only and cannot be used to diagnose, treat or manage patients.