Other search tools

About this data

The publication data currently available has been vetted by Vanderbilt faculty, staff, administrators and trainees. The data itself is retrieved directly from NCBI's PubMed and is automatically updated on a weekly basis to ensure accuracy and completeness.

If you have any questions or comments, please contact us.

Results: 1 to 10 of 213

Publication Record

Connections

Generation of MLC-2v-tdTomato knock-in reporter mouse line.
Zhang Z, Nam YJ
(2018) Genesis 56: e23256
MeSH Terms: Animals, Gene Knock-In Techniques, Genes, Reporter, Lycopersicon esculentum, Mice, Mice, Inbred C57BL, Mice, Inbred Strains, Mice, Transgenic, Myosin Light Chains
Show Abstract · Added April 2, 2019
MLC-2v is a myosin light chain regulatory protein which is specifically expressed in ventricular cardiomyocytes and slow twitch skeletal muscle cells. MLC-2v plays critical roles in ventricular maturation during heart development. Mice lacking MLC-2v are embryonic lethal due to heart failure associated with abnormal myofibrillar organization of ventricular cardiomyocytes. To study the development of ventricular cardiac muscle and slow twitch skeletal muscle, we generated a new MLC-2v reporter mouse line by knocking-in a tdTomato reporter cassette into 3' UTR of the MLC-2v gene without disrupting the endogenous gene. Our results demonstrated specific MLC-2v-tdTomato knock-in reporter expression in ventricular cardiomyocytes and slow twitch muscle during myogenesis, precisely recapitulating the spatiotemporal expression pattern of endogenous MLC-2v. No tdTomato expression was observed in the atria, fast twitch muscle or other organs throughout development into adulthood. Isolated neonatal and adult ventricular cardiomyocytes uniformly express tdTomato. Taken together, MLC-2v-tdTomato knock-in reporter mouse model described in this article will serve as a valuable tool to study cardiac chamber and skeletal muscle specification during development and regeneration by overcoming the pitfalls of transgenic strategies.
© 2018 Wiley Periodicals, Inc.
0 Communities
1 Members
0 Resources
MeSH Terms
CRISPR/Cas9 engineering of a KIM-1 reporter human proximal tubule cell line.
Veach RA, Wilson MH
(2018) PLoS One 13: e0204487
MeSH Terms: Acute Kidney Injury, CRISPR-Cas Systems, Cell Line, Cisplatin, Gene Knock-In Techniques, Gene Targeting, Genes, Reporter, Genetic Engineering, Glucose, Green Fluorescent Proteins, Hepatitis A Virus Cellular Receptor 1, Homologous Recombination, Humans, Kidney Tubules, Proximal, Luciferases, Up-Regulation
Show Abstract · Added December 13, 2018
We used the CRISPR/Cas9 system to knock-in reporter transgenes at the kidney injury molecule-1 (KIM-1) locus and isolated human proximal tubule cell (HK-2) clones. PCR verified targeted knock-in of the luciferase and eGFP reporter at the KIM-1 locus. HK-2-KIM-1 reporter cells responded to various stimuli including hypoxia, cisplatin, and high glucose, indicative of upregulation of KIM-1 expression. We attempted using CRISPR/Cas9 to also engineer the KIM-1 reporter in telomerase-immortalized human RPTEC cells. However, these cells demonstrated an inability to undergo homologous recombination at the target locus. KIM-1-reporter human proximal tubular cells could be valuable tools in drug discovery for molecules inhibiting kidney injury. Additionally, our gene targeting strategy could be used in other cell lines to evaluate the biology of KIM-1 in vitro or in vivo.
0 Communities
1 Members
0 Resources
16 MeSH Terms
Single-Cell Transcriptomic Profiling of Pluripotent Stem Cell-Derived SCGB3A2+ Airway Epithelium.
McCauley KB, Alysandratos KD, Jacob A, Hawkins F, Caballero IS, Vedaie M, Yang W, Slovik KJ, Morley M, Carraro G, Kook S, Guttentag SH, Stripp BR, Morrisey EE, Kotton DN
(2018) Stem Cell Reports 10: 1579-1595
MeSH Terms: Animals, Cell Differentiation, Cell Line, Cell Lineage, Cell Plasticity, Epithelium, Gene Expression Profiling, Genes, Reporter, Humans, Induced Pluripotent Stem Cells, Kinetics, Lung, Mice, Secretoglobins, Sequence Analysis, RNA, Single-Cell Analysis, Solubility, Spheroids, Cellular, Time Factors, Transcriptome, Wnt Signaling Pathway
Show Abstract · Added April 1, 2019
Lung epithelial lineages have been difficult to maintain in pure form in vitro, and lineage-specific reporters have proven invaluable for monitoring their emergence from cultured pluripotent stem cells (PSCs). However, reporter constructs for tracking proximal airway lineages generated from PSCs have not been previously available, limiting the characterization of these cells. Here, we engineer mouse and human PSC lines carrying airway secretory lineage reporters that facilitate the tracking, purification, and profiling of this lung subtype. Through bulk and single-cell-based global transcriptomic profiling, we find PSC-derived airway secretory cells are susceptible to phenotypic plasticity exemplified by the tendency to co-express both a proximal airway secretory program as well as an alveolar type 2 cell program, which can be minimized by inhibiting endogenous Wnt signaling. Our results provide global profiles of engineered lung cell fates, a guide for improving their directed differentiation, and a human model of the developing airway.
Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.
0 Communities
1 Members
0 Resources
21 MeSH Terms
Differentiation of Human Pluripotent Stem Cells into Functional Lung Alveolar Epithelial Cells.
Jacob A, Morley M, Hawkins F, McCauley KB, Jean JC, Heins H, Na CL, Weaver TE, Vedaie M, Hurley K, Hinds A, Russo SJ, Kook S, Zacharias W, Ochs M, Traber K, Quinton LJ, Crane A, Davis BR, White FV, Wambach J, Whitsett JA, Cole FS, Morrisey EE, Guttentag SH, Beers MF, Kotton DN
(2017) Cell Stem Cell 21: 472-488.e10
MeSH Terms: Base Sequence, Cell Differentiation, Cell Line, Cell Proliferation, Cell Self Renewal, Cell Separation, Epithelial Cells, Gene Expression Profiling, Genes, Reporter, Humans, Lung Diseases, Models, Biological, Pluripotent Stem Cells, Pulmonary Alveoli, Pulmonary Surfactants, Thyroid Nuclear Factor 1, Time Factors, Wnt Proteins, Wnt Signaling Pathway
Show Abstract · Added April 1, 2019
Lung alveoli, which are unique to air-breathing organisms, have been challenging to generate from pluripotent stem cells (PSCs) in part because there are limited model systems available to provide the necessary developmental roadmaps for in vitro differentiation. Here we report the generation of alveolar epithelial type 2 cells (AEC2s), the facultative progenitors of lung alveoli, from human PSCs. Using multicolored fluorescent reporter lines, we track and purify human SFTPC+ alveolar progenitors as they emerge from endodermal precursors in response to stimulation of Wnt and FGF signaling. Purified PSC-derived SFTPC+ cells form monolayered epithelial "alveolospheres" in 3D cultures without the need for mesenchymal support, exhibit self-renewal capacity, and display additional AEC2 functional capacities. Footprint-free CRISPR-based gene correction of PSCs derived from patients carrying a homozygous surfactant mutation (SFTPB) restores surfactant processing in AEC2s. Thus, PSC-derived AEC2s provide a platform for disease modeling and future functional regeneration of the distal lung.
Copyright © 2017 Elsevier Inc. All rights reserved.
0 Communities
1 Members
0 Resources
MeSH Terms
Lef1-dependent hypothalamic neurogenesis inhibits anxiety.
Xie Y, Kaufmann D, Moulton MJ, Panahi S, Gaynes JA, Watters HN, Zhou D, Xue HH, Fung CM, Levine EM, Letsou A, Brennan KC, Dorsky RI
(2017) PLoS Biol 15: e2002257
MeSH Terms: Animals, Anxiety, Behavior, Animal, Biomarkers, Drosophila Proteins, Drosophila melanogaster, Female, Gene Expression Regulation, Genes, Reporter, Humans, Hypothalamus, Lymphoid Enhancer-Binding Factor 1, Male, Mice, Knockout, Mice, Transgenic, Mutation, Nerve Tissue Proteins, Neurogenesis, Neurons, Species Specificity, Transcription Factors, Zebrafish, Zebrafish Proteins
Show Abstract · Added February 14, 2018
While innate behaviors are conserved throughout the animal kingdom, it is unknown whether common signaling pathways regulate the development of neuronal populations mediating these behaviors in diverse organisms. Here, we demonstrate that the Wnt/ß-catenin effector Lef1 is required for the differentiation of anxiolytic hypothalamic neurons in zebrafish and mice, although the identity of Lef1-dependent genes and neurons differ between these 2 species. We further show that zebrafish and Drosophila have common Lef1-dependent gene expression in their respective neuroendocrine organs, consistent with a conserved pathway that has diverged in the mouse. Finally, orthologs of Lef1-dependent genes from both zebrafish and mouse show highly correlated hypothalamic expression in marmosets and humans, suggesting co-regulation of 2 parallel anxiolytic pathways in primates. These findings demonstrate that during evolution, a transcription factor can act through multiple mechanisms to generate a common behavioral output, and that Lef1 regulates circuit development that is fundamentally important for mediating anxiety in a wide variety of animal species.
0 Communities
1 Members
0 Resources
23 MeSH Terms
Principles of Broad and Potent Antiviral Human Antibodies: Insights for Vaccine Design.
Crowe JE
(2017) Cell Host Microbe 22: 193-206
MeSH Terms: Animals, Antibodies, Monoclonal, Antibodies, Neutralizing, Antibodies, Viral, Antigens, Viral, Antiviral Agents, Cross Reactions, Drug Design, Genes, Reporter, Humans, Immunity, Models, Molecular, Neutralization Tests, Protein Structure, Quaternary, Vaccination, Vaccines, Viral Envelope Proteins, Virus Diseases
Show Abstract · Added March 14, 2018
Antibodies are the principal immune effectors that mediate protection against reinfection following viral infection or vaccination. Robust techniques for human mAb isolation have been developed in the last decade. The study of human mAbs isolated from subjects with prior immunity has become a mainstay for rational structure-based, next-generation vaccine development. The plethora of detailed molecular and genetic studies coupling the structure of antigen-antibody complexes with their antiviral function has begun to reveal common principles of critical interactions on which we can build better vaccines and therapeutic antibodies. This review outlines the approaches to isolating and studying human antiviral mAbs and discusses the common principles underlying the basis for their activity. This review also examines progress toward the goal of achieving a comprehensive understanding of the chemical and physical basis for molecular recognition of viral surface proteins in order to build predictive molecular models that can be used for vaccine design.
Copyright © 2017. Published by Elsevier Inc.
0 Communities
1 Members
0 Resources
18 MeSH Terms
A Chimeric Egfr Protein Reporter Mouse Reveals Egfr Localization and Trafficking In Vivo.
Yang YP, Ma H, Starchenko A, Huh WJ, Li W, Hickman FE, Zhang Q, Franklin JL, Mortlock DP, Fuhrmann S, Carter BD, Ihrie RA, Coffey RJ
(2017) Cell Rep 19: 1257-1267
MeSH Terms: Adult Stem Cells, Amphiregulin, Animals, Embryo, Mammalian, ErbB Receptors, Genes, Reporter, Green Fluorescent Proteins, Hepatocytes, Intestinal Mucosa, Mice, Microscopy, Fluorescence, Protein Transport, Recombinant Proteins, Transgenes
Show Abstract · Added June 21, 2017
EGF receptor (EGFR) is a critical signaling node throughout life. However, it has not been possible to directly visualize endogenous Egfr in mice. Using CRISPR/Cas9 genome editing, we appended a fluorescent reporter to the C terminus of the Egfr. Homozygous reporter mice appear normal and EGFR signaling is intact in vitro and in vivo. We detect distinct patterns of Egfr expression in progenitor and differentiated compartments in embryonic and adult mice. Systemic delivery of EGF or amphiregulin results in markedly different patterns of Egfr internalization and trafficking in hepatocytes. In the normal intestine, Egfr localizes to the crypt rather than villus compartment, expression is higher in adjacent epithelium than in intestinal tumors, and following colonic injury expression appears in distinct cell populations in the stroma. This reporter, under control of its endogenous regulatory elements, enables in vivo monitoring of the dynamics of Egfr localization and trafficking in normal and disease states.
Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.
1 Communities
4 Members
2 Resources
14 MeSH Terms
Kidney-specific transposon-mediated gene transfer in vivo.
Woodard LE, Cheng J, Welch RC, Williams FM, Luo W, Gewin LS, Wilson MH
(2017) Sci Rep 7: 44904
MeSH Terms: Acute Kidney Injury, Animals, DNA Transposable Elements, Erythropoietin, Gene Expression, Gene Expression Regulation, Gene Transfer Techniques, Genes, Reporter, Genetic Vectors, Hydrodynamics, Immunosuppressive Agents, Kidney, Male, Mice, Organ Specificity, Promoter Regions, Genetic, Transfection
Show Abstract · Added September 11, 2017
Methods enabling kidney-specific gene transfer in adult mice are needed to develop new therapies for kidney disease. We attempted kidney-specific gene transfer following hydrodynamic tail vein injection using the kidney-specific podocin and gamma-glutamyl transferase promoters, but found expression primarily in the liver. In order to achieve kidney-specific transgene expression, we tested direct hydrodynamic injection of a DNA solution into the renal pelvis and found that luciferase expression was strong in the kidney and absent from extra-renal tissues. We observed heterogeneous, low-level transfection of the collecting duct, proximal tubule, distal tubule, interstitial cells, and rarely glomerular cells following injection. To assess renal injury, we performed the renal pelvis injections on uninephrectomised mice and found that their blood urea nitrogen was elevated at two days post-transfer but resolved within two weeks. Although luciferase expression quickly decreased following renal pelvis injection, the use of the piggyBac transposon system improved long-term expression. Immunosuppression with cyclophosphamide stabilised luciferase expression, suggesting immune clearance of the transfected cells occurs in immunocompetent animals. Injection of a transposon expressing erythropoietin raised the haematocrit, indicating that the developed injection technique can elicit a biologic effect in vivo. Hydrodynamic renal pelvis injection enables transposon mediated-kidney specific gene transfer in adult mice.
0 Communities
3 Members
1 Resources
17 MeSH Terms
Comparative genetic screens in human cells reveal new regulatory mechanisms in WNT signaling.
Lebensohn AM, Dubey R, Neitzel LR, Tacchelly-Benites O, Yang E, Marceau CD, Davis EM, Patel BB, Bahrami-Nejad Z, Travaglini KJ, Ahmed Y, Lee E, Carette JE, Rohatgi R
(2016) Elife 5:
MeSH Terms: Casein Kinase I, Cytoskeletal Proteins, Gene Expression Regulation, Gene Regulatory Networks, Genes, Reporter, Genetic Testing, Haploidy, Humans, Wnt Proteins, Wnt Signaling Pathway
Show Abstract · Added February 13, 2017
The comprehensive understanding of cellular signaling pathways remains a challenge due to multiple layers of regulation that may become evident only when the pathway is probed at different levels or critical nodes are eliminated. To discover regulatory mechanisms in canonical WNT signaling, we conducted a systematic forward genetic analysis through reporter-based screens in haploid human cells. Comparison of screens for negative, attenuating and positive regulators of WNT signaling, mediators of R-spondin-dependent signaling and suppressors of constitutive signaling induced by loss of the tumor suppressor adenomatous polyposis coli or casein kinase 1α uncovered new regulatory features at most levels of the pathway. These include a requirement for the transcription factor AP-4, a role for the DAX domain of AXIN2 in controlling β-catenin transcriptional activity, a contribution of glycophosphatidylinositol anchor biosynthesis and glypicans to R-spondin-potentiated WNT signaling, and two different mechanisms that regulate signaling when distinct components of the β-catenin destruction complex are lost. The conceptual and methodological framework we describe should enable the comprehensive understanding of other signaling systems.
0 Communities
1 Members
0 Resources
10 MeSH Terms
Temporal self-regulation of transposition through host-independent transposase rodlet formation.
Woodard LE, Downes LM, Lee YC, Kaja A, Terefe ES, Wilson MH
(2017) Nucleic Acids Res 45: 353-366
MeSH Terms: Animals, DNA Transposable Elements, Female, Gene Expression Regulation, Genes, Reporter, HEK293 Cells, HeLa Cells, Humans, Insect Proteins, Luciferases, Male, Mice, Optical Imaging, Time-Lapse Imaging, Transposases, Tribolium
Show Abstract · Added December 8, 2017
Transposons are highly abundant in eukaryotic genomes, but their mobilization must be finely tuned to maintain host organism fitness and allow for transposon propagation. Forty percent of the human genome is comprised of transposable element sequences, and the most abundant cut-and-paste transposons are from the hAT superfamily. We found that the hAT transposase TcBuster from Tribolium castaneum formed filamentous structures, or rodlets, in human tissue culture cells, after gene transfer to adult mice, and ex vivo in cell-free conditions, indicating that host co-factors or cellular structures were not required for rodlet formation. Time-lapsed imaging of GFP-laced rodlets in human cells revealed that they formed quickly in a dynamic process involving fusion and fission. We delayed the availability of the transposon DNA and found that transposition declined after transposase concentrations became high enough for visible transposase rodlets to appear. In combination with earlier findings for maize Ac elements, these results give insight into transposase overproduction inhibition by demonstrating that the appearance of transposase protein structures and the end of active transposition are simultaneous, an effect with implications for genetic engineering and horizontal gene transfer.
Published by Oxford University Press on behalf of Nucleic Acids Research 2016. This work is written by (a) US Government employee(s) and is in the public domain in the US.
0 Communities
2 Members
0 Resources
16 MeSH Terms