A5074994344- Animal Genetics and Reproduction
- Reproductive Biology and Fertility
- Pluripotent Stem Cells Research
- CRISPR and Genetic Engineering
- Virus-based gene therapy research
- Xenotransplantation and immune response
- Viral gastroenteritis research and epidemiology
- Animal Virus Infections Studies
- Molecular Biology Techniques and Applications
- Renal and related cancers
- Tissue Engineering and Regenerative Medicine
- Genetic and phenotypic traits in livestock
- Epigenetics and DNA Methylation
- Reproductive Physiology in Livestock
- Genetic Mapping and Diversity in Plants and Animals
- Pediatric health and respiratory diseases
- Neonatal Respiratory Health Research
- Birth, Development, and Health
- Transplantation: Methods and Outcomes
- Reproductive System and Pregnancy
- Viral Infectious Diseases and Gene Expression in Insects
- Cancer Genomics and Diagnostics
- Infant Nutrition and Health
- Cystic Fibrosis Research Advances
- RNA Interference and Gene Delivery
University of Missouri
2016-2025
Women's Resource Center
2024
The Graduate Center, CUNY
2016-2018
University of Iowa
2008
Howard Hughes Medical Institute
2008
Almost two decades after CFTR was identified as the gene responsible for cystic fibrosis (CF), we still lack answers to many questions about pathogenesis of disease, and it remains incurable. Mice with a disrupted have greatly facilitated CF studies, but mutant mice do not develop characteristic manifestations human CF, including abnormalities pancreas, lung, intestine, liver, other organs. Because pigs share anatomical physiological features humans, generated targeted disruption both...
Targeted modification of the pig genome can be challenging. Recent applications CRISPR/Cas9 system hold promise for improving efficacy editing. When a designed targeting CD163 or CD1D was introduced into somatic cells, it highly efficient in inducing mutations. these mutated cells were used with cell nuclear transfer, offspring modifications created. delivered vitro produced presumptive porcine zygotes, effective creating mutations eGFP, CD163, and (100% efficiency blastocyst stage embryos);...
Progress toward understanding the pathogenesis of cystic fibrosis (CF) and developing effective therapies has been hampered by lack a relevant animal model. CF mice fail to develop lung pancreatic disease that cause most morbidity mortality in patients with CF. Pigs may be better animals than which model human genetic diseases because their anatomy, biochemistry, physiology, size, genetics are more similar those humans. However, date, gene-targeted mammalian models have not reported for any...
The alphacoronaviruses, transmissible gastroenteritis virus (TGEV) and Porcine epidemic diarrhea (PEDV) are sources of high morbidity mortality in neonatal pigs, a consequence dehydration caused by the infection necrosis enterocytes. biological relevance amino peptidase N (ANPEP) as putative receptor for TGEV PEDV pigs was evaluated using CRISPR/Cas9 to edit exon 2 ANPEP resulting premature stop codon. Knockout possessing null phenotype age matched wild type were challenged with either or...
ABSTRACT Surface expression of SIGLEC1, also known as sialoadhesin or CD169, is considered a primary determinant the permissiveness porcine alveolar macrophages for infection by reproductive and respiratory syndrome virus (PRRSV). In vitro , attachment internalization PRRSV are dependent on interaction between sialic acid virion surface binding domain SIGLEC1 gene. To test role in infection, gene knockout pig was created removing part exon 1 all exons 2 3 The resulting ablated but had no...
CD163 knockout (KO) pigs are resistant to infection with genotype 2 (type 2) porcine reproductive and respiratory syndrome virus (PRRSV). Furthermore, the substitution of scavenger receptor cysteine-rich (SRCR) domain 5 a homolog human CD163-like (hCD163L1) SRCR 8 confers resistance transfected HEK cells type 1 PRRSV. As means understand role in PRRSV both viruses, were genetically modified (GM) possess one following genotypes: complete CD163, deletions within 5, or replacement (domain swap)...
Differential mRNA expression patterns were evaluated between germinal vesicle oocytes (pgvo), four-cell (p4civv), blastocyst (pblivv), and in vitro-produced (p4civp) (pblivp) stage embryos to determine key transcripts responsible for early embryonic development the pig. Five comparisons made: pgvo p4civv, p4civv pblivv, p4civp, pblivv pblivp. ANOVA (P < 0.05) was performed with Benjamini Hochberg false-discovery-rate multiple correction test on each comparison. A comparison of resulted 3214,...
Abstract Fertile offspring from somatic cell nuclear transfer (SCNT) is the goal of most cloning laboratories. For this process to be successful, a number events must occur correctly. First donor nucleus in state that amenable remodeling and subsequent genomic reprogramming. The introduced into an oocyte cytoplasm capable facilitating remodeling. then adequately stimulated initiate development. Finally resulting embryo cultured environment compatible with development particular embryo. Much...
Glucose metabolism in preimplantation embryos has traditionally been viewed from a somatic cell viewpoint. Here, we show that gene expression early is similar to rapidly dividing cancer cells. In vitro-produced pig blastocysts were subjected deep-sequencing, and found express two variants have ascribed importance (HK2 the M2 variant of PKM2). Development was monitored quantified additional cultured low or high O(2) (5% CO(2), 5% O(2), 90% N(2) vs. CO(2) air). blastocyst stage atmospheres...
Abstract The proposed signal for maternal recognition of pregnancy in pigs is estrogen (E2), produced by the elongating conceptuses between days 11 to 12 with a more sustained increase during conceptus attachment and placental development on 15 30. To understand role E2 porcine elongation establishment, loss-of-function study was conducted editing aromatase (CYP19A1) using CRISPR/Cas9 technology. Wild-type (CYP19A1+/+) (CYP19A1−/−) fibroblast cells were used create embryos through somatic...
Nuclear transfer efficiency in the pig is low and thought to be caused by inadequate nuclear reprogramming. The objective of this study was identify differentially represented transcripts vivo derived (BLIVV), vitro fertilized (BLIVF), or (NT, three different activation methods) blastocyst stage embryos donor cell line microarray analysis, determine if treatment reconstructed with histone deacetylase inhibitor, Scriptaid (NTS), for 14 h postactivation would correct gene expression a subset...
In vitro embryo culture systems promote development at rates lower than in vivo systems. The goal of this project was to discover transcripts that may be responsible for a decrease competency blastocyst-stage embryos cultured vitro. Gilts were artificially inseminated on the first day estrus, and Day 2, one oviduct tip uterine horn flushed recovered porcine zygote medium 3 4 days. On 6, gilts euthanized contralateral obtain derived embryos. Total RNA extracted from three pools 10 blastocysts...
Evidence in many species has suggested that those embryos cleave earliest after fertilization are more developmentally competent than relatively later fertilization. Herein we document this phenomenon porcine vitro-fertilized (IVF), somatic cell nuclear transfer (SCNT), and parthenogenetic (PA) embryos. In vitro-matured pig oocytes were used to generate IVF, SCNT, PA At 24 hr post-activation (or insemination; hpa/hpi), visually assessed, cleaved moved into a new culture well. This process...
The coronaviruses, porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis (TGEV), and deltacoronavirus (PDCoV) represent important sources of neonatal on pig farms. requirement for aminopeptidase N (APN) as a receptor TGEV, but not PEDV, is well established. In this study, the biological relevance APN PDCoV was tested by using CRISPR/Cas9 to knockout gene, ANPEP, in pigs. Porcine alveolar macrophages (PAMs) from ANPEP (KO) pigs showed resistance infection. However, lung...
Pigs are playing an increasingly vital role as translational biomedical models for studying human pathophysiology. The annotation of the pig genome was a huge step forward in translatability pigs model various diseases. Similarities between humans and terms anatomy, physiology, genetics, immunology have allowed to become comprehensive preclinical With diverse range, from craniofacial ophthalmology reproduction, wound healing, musculoskeletal, cancer, provided seminal understanding This...
Abstract The following experiments compared the efficiency of three fusion/activation protocols somatic cell nuclear transfer (SCNT) with porcine cells transfected enhanced green fluorescent protein driven by chicken β‐actin/rabbit β‐globin hybrid promoter (pCAGG‐EGFP). included electrical (NT1), followed treatment a reversible proteasomal inhibitor MG132 (NT2) and fusion in low Ca 2+ chemical activation thimerosal/dithiothreitol (NT3). Data were collected at Days 6, 12, 14, 30, 114...
KDM5B (JARID1B/PLU1) is a H3K4me2/3 histone demethylase that implicated in cancer development and proliferation also indispensable for embryonic stem cell self-renewal, fate, murine development. However, little known about the role of during preimplantation embryo Here we show critical to porcine was found be expressed stage-specific manner, consistent with demethylation H3K4me3, highest expression being observed from 4-cell blastocyst stages. Knockdown by morpholino antisense...
Most in vitro culture conditions are less-than-optimal for embryo development. Here, we used a transcriptional-profiling database to identify culture-induced differences gene expression porcine blastocysts compared vivo-produced counterparts. Genes involved glycine transport (SLC6A9), metabolism (GLDC, GCSH, DLD, and AMT), serine (PSAT1, PSPH, PHGDH) were differentially expressed. Addition of 10 mM the medium (currently containing 0.1 mM) reduced abundance SLC6A9 transcript increased total...
Submucosal glands (SMGs) are a prominent structure that lines human cartilaginous airways. Although it has been assumed SMGs contribute to respiratory defense, hypothesis gone without direct test. Therefore, we studied pigs, which have lungs like humans, and disrupted the gene for ectodysplasin ( EDA-KO ), initiates SMG development. pigs lacked throughout Their airway surface liquid had reduced ability kill bacteria, consistent with production of antimicrobials. In wild-type secrete mucus...
Background Modeling cardiovascular diseases in mice has provided invaluable insights into the cause of congenital heart disease. However, small size mouse precluded translational studies. Given current high‐efficiency gene editing, disease modeling other species is possible. The pig advantageous given its cardiac anatomy, physiology, and are similar to human infants. We profiled development generated genetically edited pigs with defects. Methods Results Pig conceptuses fetuses were collected...