SPC-Cre-ERT2 Transgenic Mouse for Temporal Gene Deletion in Alveolar Epithelial Cells

Gui, Yao-Song; Wang, Lianmei; Tian, Xinlun; Feng, Ru; Ma, Anqi; Cai, Baiqiang; Zhang, Hongbing; Xu, Kai‐Feng

doi:10.1371/journal.pone.0046076

Cited by 16 publications

(16 citation statements)

References 16 publications

(33 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Ssrp1 fl/fl mice were crossed with homozygous Rosa26 CreER T2 (CreER T2+/+ ) mice. Since the literature suggests that even one allele of CreER T2 is sufficient for the excision of a gene of interest [ 34 – 36 ], heterozygous mice for both alleles ( Ssrp1 fl /+ CreER T2+/- ) were bred to get mice homozygous for floxed- Ssrp1 with at least one allele of CreER T2 ( Ssrp1 fl/fl CreER T2+/- ). Fibroblasts were isolated from tail tissue of Ssrp1 fl/fl CreER T2+/- mice and then immortalized and transformed.…”

Section: Resultsmentioning

confidence: 99%

Uncovering the fine print of the CreERT2-LoxP system while generating a conditional knockout mouse model of Ssrp1 gene

et al. 2018

View full text Add to dashboard Cite

FAcilitates Chromatin Transcription (FACT) is a complex of SSRP1 and SPT16 that is involved in chromatin remodeling during transcription, replication, and DNA repair. FACT has been mostly studied in cell-free or single cell model systems because general FACT knockout (KO) is embryonically lethal (E3.5). FACT levels are limited to the early stages of development and stem cell niches of adult tissues. FACT is upregulated in poorly differentiated aggressive tumors. Importantly, FACT inhibition (RNAi) is lethal for tumors but not normal cells, making FACT a lucrative target for anticancer therapy. To develop a better understanding of FACT function in the context of the mammalian organism under normal physiological conditions and in disease, we aimed to generate a conditional FACT KO mouse model. Because SPT16 stability is dependent on the SSRP1-SPT16 association and the presence of SSRP1 mRNA, we targeted the Ssrp1 gene using a CreERT2- LoxP approach to generate the FACT KO model. Here, we highlight the limitations of the CreERT2-LoxP (Rosa26) system that we encountered during the generation of this model. In vitro studies showed an inefficient excision rate of ectopically expressed CreERT2 (retroviral CreERT2) in fibroblasts with homozygous floxed Ssrp1. In vitro and in vivo studies showed that the excision efficiency could only be increased with germline expression of two alleles of Rosa26CreERT2. The expression of one germline Rosa26CreERT2 allele led to the incomplete excision of Ssrp1. The limited efficiency of the CreERT2-LoxP system may be sufficient for studies involving the deletion of genes that interfere with cell growth or viability due to the positive selection of the phenotype. However, it may not be sufficient for studies that involve the deletion of genes supporting growth, or those crucial for development. Although CreERT2-LoxP is broadly used, it has limitations that have not been widely discussed. This paper aims to encourage such discussions.

show abstract

Section: Resultsmentioning

confidence: 99%

Uncovering the fine print of the CreERT2-LoxP system while generating a conditional knockout mouse model of Ssrp1 gene

et al. 2018

View full text Add to dashboard Cite

show abstract

“…Further studies crossing the Il17ra fl/fl mice with additional epithelial specific CRE lines are needed to determine the exact roles of epithelial IL-17R in pulmonary host defense as well as chronic lung inflammation in other cell lineages. For example, a recently established mouse model incorporated a tamoxifen-inducible Cre recombinase (Cre-ERT2) under the control of the human surfactant protein C (SPC) promoter which deletes in distal lung epithelium (Gui et al, 2012). Crossing this line with the IL-17R flox mice would permit ablation of IL-17R in the distal lung in a temporally controlled manner.…”

Section: Discussionmentioning

confidence: 99%

IL-17 Receptor Signaling in the Lung Epithelium Is Required for Mucosal Chemokine Gradients and Pulmonary Host Defense against K. pneumoniae

Chen

Eddens

Trevejo-Nuñez

et al. 2016

Cell Host & Microbe

117

108

View full text Add to dashboard Cite

Summary The cytokine IL17, and signaling via its heterodimeric IL-17RA/IL-17RC receptor, is critical for host defense against extracellular bacterial and fungal pathogens. Polarized lung epithelial cells express IL-17RA and IL-17RC basolaterally. However, their contribution to IL-17-dependent pulmonary defenses in vivo remains to be determined. To address this, we generated mice with conditional deletion of Il17ra or Il17rc in Scgb1a1-expressing club cells, a major component of the murine bronchiolar epithelium. These mice displayed an impaired ability to recruit neutrophils into the airway lumen in response to IL17, a defect in bacterial clearance upon mucosal challenge with the pulmonary pathogen Klebsiella pneumoniae, and substantially reduced epithelial expression of the chemokine Cxcl5. Neutrophil recruitment and bacterial clearance were restored by intranasal administration of recombinant CXCL5. Our data show that IL-17R signaling in the lung epithelium plays a critical role in establishing chemokine gradients that are essential for mucosal immunity against pulmonary bacterial pathogens.

show abstract

“…A number of in vivo and in vitro models are available to aid in these studies. Cell-type-specific transgenic mice targeting AECII (56) or AECI (57) may be used to…”

Section: Potential Future Approachesmentioning

confidence: 99%

Alveolar Epithelial Cell–Derived Mediators: Potential Direct Regulators of Large Airway and Vascular Responses

Oczypok

Perkins

Oury

2017

Am J Respir Cell Mol Biol

View full text Add to dashboard Cite

Bronchial epithelial cells and pulmonary endothelial cells are thought to be the primary modulators of conducting airways and vessels, respectively. However, histological examination of both mouse and human lung tissue reveals that alveolar epithelial cells (AECs) line the adventitia of large airways and vessels and thus are also in a position to directly regulate these structures. The primary purpose of this perspective is to highlight the fact that AECs coat the adventitial surface of every vessel and airway in the lung parenchyma. This localization is ideal for transmitting signals that can contribute to physiologic and pathologic responses in vessels and airways. A few examples of mediators produced by AECs that may contribute to vascular and airway responses are provided to illustrate some of the potential effects that AECs may modulate.

show abstract

SPC-Cre-ERT2 Transgenic Mouse for Temporal Gene Deletion in Alveolar Epithelial Cells

Cited by 16 publications

References 16 publications

Uncovering the fine print of the CreERT2-LoxP system while generating a conditional knockout mouse model of Ssrp1 gene

Uncovering the fine print of the CreERT2-LoxP system while generating a conditional knockout mouse model of Ssrp1 gene

IL-17 Receptor Signaling in the Lung Epithelium Is Required for Mucosal Chemokine Gradients and Pulmonary Host Defense against K. pneumoniae

Alveolar Epithelial Cell–Derived Mediators: Potential Direct Regulators of Large Airway and Vascular Responses

Contact Info

Product

Resources

About