Transgenic rescue of GATA-1–deficient mice with GATA-1 lacking a FOG-1 association site phenocopies patients with X-linked thrombocytopenia

Blood ◽  
2004 ◽  
Vol 103 (7) ◽  
pp. 2560-2567 ◽  
Author(s):  
Ritsuko Shimizu ◽  
Kinuko Ohneda ◽  
James Douglas Engel ◽  
Cecelia D. Trainor ◽  
Masayuki Yamamoto
Keyword(s):  
Transgene Expression ◽  
Germ Line ◽  
Embryonic Lethality ◽  
Regulatory Domain ◽  
Mouse Development ◽  
Transgenic Expression ◽  
Stress Erythropoiesis ◽  
Unique Model ◽  
Cytoplasmic Maturation ◽  
Deficient Mice

Abstract Association of GATA-1 and its cofactor Friend of GATA-1 (FOG-1) is essential for erythroid and megakaryocyte development. To assess functions of GATA-1–FOG-1 association during mouse development, we used the GATA-1 hematopoietic regulatory domain to generate transgenic mouse lines expressing a mutant GATA-1, which contains a substitution of glycine 205 for valine (V205G) that abrogates its association with FOG-1. We examined whether the transgenic expression of mutant GATA-1 rescues GATA-1 germ line mutants from embryonic lethality. In high-expressor lines we observed that the GATA-1V205G rescues GATA-1–deficient mice from embryonic lethality at the expected frequency, revealing that excess GATA-1V205G can eliminate the lethal anemia that is due to GATA-1 deficiency. In contrast, transgene expression comparable to the endogenous GATA-1 level resulted in much lower frequency of rescue, indicating that the GATA-1–FOG-1 association is critical for normal embryonic hematopoiesis. Rescued mice in these analyses exhibit thrombocytopenia and display dysregulated proliferation and impaired cytoplasmic maturation of megakaryocytes. Although anemia is not observed under steady-state conditions, stress erythropoiesis is attenuated in the rescued mice. Our findings reveal an indispensable role for the association of GATA-1 and FOG-1 during late-stage megakaryopoiesis and provide a unique model for X-linked thrombocytopenia with inherited GATA-1 mutation.

Rescue of Prothrombin-deficiency by Transgene Expression in Mice

2002 ◽  
Vol 88 (12) ◽  
pp. 984-991 ◽  
Author(s):  
William Sun ◽  
Mallory Coleman ◽  
David Witte ◽  
Sandra Degen
Keyword(s):  
Transgenic Mice ◽  
Transgene Expression ◽  
Embryonic Lethality ◽  
The Other ◽  
Model Systems ◽  
Bleeding Events ◽  
Specific Expression ◽  
Spontaneous Bleeding ◽  
Knock Out ◽  
Transgenic Expression

SummaryProthrombin has diverse biological functions in addition to its well established role in blood coagulation. In order to study these functions in more detail mouse model systems are needed. Since deficiency of prothrombin in mice results in partial embryonic lethality and neonatal death, alternative approaches are required to study the biology of prothrombin in the adult mouse. The liver is the major site of synthesis of prothrombin and therefore liver-specific promoters were used to express prothrombin in transgenic mice. Mice generated from crosses with these transgenic mice and mice hemizygous for the knock-out allele were used to test whether liver-specific expression is sufficient to correct the phenotype of null mice and whether liver-specific expression is sufficient for the development and survival of mice to adulthood. The mouse albumin promoter/enhancer was used initially for transgene expression without success in obtaining transgene positive, endogenous prothrombin null mice. Two lines of transgene positive, endogenous prothrombin deficient mice were obtained using the mouse transthyretin (TTR) promoter/enhancer driving expression of a human prothrombin cDNA. One line was able to rescue both the embryonic and the neonatal lethality while the other line was only able to correct the embryonic lethality. Expression of prothrombin was restricted to the liver and stomach in one line and to the liver, pancreas, stomach and kidney in the other line of mice. Thrombin activity for one line was determined to be at 5-10% of wildtype levels. These mice developed normally and did not have spontaneous bleeding events unless traumatized. Therefore, transgenic expression of human prothrombin is sufficient for the rescue of the lethality found for prothrombin deficiency in mice.

scholarly journals Naa12 rescues embryonic lethality in Naa10-Deficient Mice in the amino-terminal acetylation pathway

2020 ◽  
Author(s):  
Hyae Yon Kweon ◽  
Mi-Ni Lee ◽  
Max Dörfel ◽  
Seungwoon Seo ◽  
Leah Gottlieb ◽  
...  
Keyword(s):  
Embryonic Lethality ◽  
Mouse Development ◽  
Post Translational Modifications ◽  
Amino Terminal ◽  
Neonatal Lethality ◽  
Enormous Amount ◽  
Deficient Mice ◽  
Urogenital Anomalies

AbstractThere is an enormous amount of variation in proteins introduced by co- and post-translational modifications, including N-terminal acetylation (NTA), catalyzed by a set of N-terminal acetyltransferases (NATs). The NatA complex (including X-linked Naa10 and Naa15) is the major acetyltransferase, with 40–50% of all mammalian proteins being potential substrates. However, the overall role of NTA on a whole-organism level is poorly understood, particularly in mammals. Male mice lacking Naa10 show no globally apparent in vivo NTA impairment and, surprisingly, do not exhibit embryonic lethality. Rather Naa10 nulls display increased neonatal lethality, and the majority of surviving undersized mutants exhibit a combination of hydrocephaly, cardiac defects, homeotic anterior transformation (including an extra thoracic rib), piebaldism and urogenital anomalies. The lack of complete embryonic lethality in Naa10-null mice is explained by the discovery of Naa12, a previously unannotated Naa10-like paralogue with NAT activity that genetically compensates for Naa10. Mice deficient for Naa12 have no apparent phenotype, except for decreased fertility, whereas mice doubly deficient for Naa10 and Naa12 display embryonic lethality, thus presenting the complete machinery for NatA-mediated NTA in mouse development.

Conditional inactivation of VEGF-A in areas of collagen2a1 expression results in embryonic lethality in the heterozygous state

Development ◽  
2000 ◽  
Vol 127 (7) ◽  
pp. 1445-1453 ◽  
Author(s):  
J.J. Haigh ◽  
H.P. Gerber ◽  
N. Ferrara ◽  
E.F. Wagner
Keyword(s):  
Bone Formation ◽  
Ischemic Cardiomyopathy ◽  
Embryonic Lethality ◽  
Limb Bud ◽  
Endochondral Bone Formation ◽  
Functional Importance ◽  
Mouse Development ◽  
Endochondral Bone ◽  
Myocardial Layers ◽  
Transgenic Lines

VEGF-A has been implicated in regulating the initial angiogenic invasion events that are essential for endochondral bone formation. VEGF-A mRNA expression was indeed found in the sclerotome of the developing somite and in the limb-bud mesenchyme at E10.5 in mouse development but declined during chondrogenesis and became upregulated in hypertrophic chondrocytes prior to angiogenic invasion. To determine the functional importance of VEGF-A expression in the developing chondrogenic tissues, VEGF-A was conditionally inactivated during early embryonic development using Collagen2a1-Cre transgenic lines. Deletion of a single VEGF-A allele in Collagen2a1-Cre-expressing cells results in embryonic lethality around E10.5. This lethality is characterized by aberrant development of the dorsal aorta and intersomitic blood vessels, along with defects in the developing endocardial and myocardial layers of the heart. A small percentage of VEGF(Flox)/+, Collagen2a1-Cre fetuses survive until E17.5, show aberrant endochondral bone formation and develop a heart phenotype resembling a dilated form of ischemic cardiomyopathy. These results provide insights into the function of VEGF-A in heart and endochondral bone formation and underscore the importance of tightly controlled levels of VEGF-A during development.

scholarly journals Circumventing Zygotic Lethality to Generate Maternal Mutants in Zebrafish

Biology ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 102
Author(s):  
De-Li Shi
Keyword(s):  
Germ Line ◽  
Gene Products ◽  
Transgenic Expression ◽  
Guide Rna ◽  
Guide Rnas ◽  
Transgenic Embryos ◽  
Mutant Phenotypes ◽  
Biallelic Mutations ◽  
Genome Activation ◽  
Maternal Gene

Maternal gene products accumulated during oogenesis are essential for supporting early developmental processes in both invertebrates and vertebrates. Therefore, understanding their regulatory functions should provide insights into the maternal control of embryogenesis. The CRISPR/Cas9 genome editing technology has provided a powerful tool for creating genetic mutations to study gene functions and developing disease models to identify new therapeutics. However, many maternal genes are also essential after zygotic genome activation; as a result, loss of their zygotic functions often leads to lethality or sterility, thus preventing the generation of maternal mutants by classical crossing between zygotic homozygous mutant adult animals. Although several approaches, such as the rescue of mutant phenotypes through an injection of the wild-type mRNA, germ-line replacement, and the generation of genetically mosaic females, have been developed to overcome this difficulty, they are often technically challenging and time-consuming or inappropriate for many genes that are essential for late developmental events or for germ-line formation. Recently, a method based on the oocyte transgenic expression of CRISPR/Cas9 and guide RNAs has been designed to eliminate maternal gene products in zebrafish. This approach introduces several tandem guide RNA expression cassettes and a GFP reporter into transgenic embryos expressing Cas9 to create biallelic mutations and inactivate genes of interest specifically in the developing oocytes. It is particularly accessible and allows for the elimination of maternal gene products in one fish generation. By further improving its efficiency, this method can be used for the systematic characterization of maternal-effect genes.

Embryonic lethality in Dear gene-deficient mice: new player in angiogenesis

2005 ◽  
Vol 23 (3) ◽  
pp. 257-268 ◽  
Author(s):  
Victoria L. M. Herrera ◽  
Lorenz R. B. Ponce ◽  
Pia D. Bagamasbad ◽  
Benjamin D. VanPelt ◽  
Tamara Didishvili ◽  
...  
Keyword(s):  
Angiotensin Ii ◽  
Embryonic Lethality ◽  
Female Rats ◽  
Ang Ii ◽  
Sprague Dawley ◽  
Angiotensin Ii Receptor ◽  
Endothelin 1 ◽  
Receptor Isoforms ◽  
Age Range ◽  
Deficient Mice

The dual endothelin-1/angiotensin II receptor (Dear) binds endothelin-1 (ET-1) and angiotensin II (ANG II) with equal affinities in the Dahl S/JRHS rat strain. To elucidate its physiological significance within the context of multiple receptor isoforms and diverse ET-1 and ANG II functions spanning blood pressure regulation, tumor proliferation, and angiogenesis, we characterized mouse Dear and Dear-deficient mice. Unlike null mutant models of ET-1, ANG II, and all other ET-1 and ANG II receptors, Dear−/− deficiency results in impaired angiogenesis, dysregulated neuroepithelial development, and embryonic lethality by embryonic day 12.5. Interestingly, mouse Dear does not bind ANG II, similar to Dahl R/JRHS rat Dear, but binds ET-1 and vascular endothelial growth factor (VEGF) signal peptide (VEGFsp) with equal affinities, suggesting a putative novel multifunction for VEGFsp and a parsimonious mechanism for coordination of VEGF-induced and Dear-mediated pathways. Consistent with its developmental angiogenic role, Dear inhibition results in decreased tumor growth in B16-F10 melanoma cell-induced subcutaneous tumor in female Dear+/−/C57BL6BC10 mice, but not in males (age 3.5 mo), and in 127Cs radiation-induced orthotopic mammary tumors in Sprague-Dawley female rats (age range 3–6.5 mo). Altogether, the data identify Dear as a new player in angiogenesis during development downstream to, and nonredundant with, VEGF-mediated pathways, as well as a putative modulator of tumor angiogenesis acting within a gender-specific paradigm.

Rescue of embryonic lethality in Mdm2-deficient mice by absence of p53

Nature ◽  
1995 ◽  
Vol 378 (6553) ◽  
pp. 206-208 ◽  
Author(s):  
Stephen N. Jones ◽  
Amy E. Roe ◽  
Lawrence A. Donehower ◽  
Allan Bradley
Keyword(s):  
Embryonic Lethality ◽  
Deficient Mice

scholarly journals A XRCC4 mutant mouse, a model for human X4 syndrome, reveals interplays with Xlf, PAXX, and ATM in lymphoid development

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Benoit Roch ◽  
Vincent Abramowski ◽  
Olivier Etienne ◽  
Stefania Musilli ◽  
Pierre David ◽  
...  
Keyword(s):  
Dna Repair ◽  
Clinical Manifestations ◽  
Adaptive Immune System ◽  
Embryonic Lethality ◽  
Dna Ligase ◽  
Mouse Development ◽  
Dna Ligase Iv ◽  
Ligase Iv ◽  
A Minor

We developed a Xrcc4M61R separation of function mouse line to overcome the embryonic lethality of Xrcc4 deficient mice. XRCC4M61R protein does not interact with Xlf, thus obliterating XRCC4-Xlf filament formation while preserving the ability to stabilize DNA Ligase IV. X4M61R mice, which are DNA repair deficient, phenocopy the Nhej1-/- (known as Xlf -/-) setting with a minor impact on the development of the adaptive immune system. The core NHEJ DNA repair factor XRCC4 is therefore not mandatory for V(D)J recombination aside from its role in stabilizing DNA ligase IV. In contrast, Xrcc4M61R mice crossed on Paxx-/-, Nhej1-/-, or Atm-/- backgrounds are severely immunocompromised, owing to aborted V(D)J recombination as in Xlf-Paxx and Xlf-Atm double KO settings. Furthermore, massive apoptosis of post-mitotic neurons causes embryonic lethality of Xrcc4M61R -Nhej1-/- double mutants. These in vivo results reveal new functional interplays between XRCC4 and PAXX, ATM and Xlf in mouse development and provide new insights in the understanding of the clinical manifestations of human XRCC4 deficient condition, in particular its absence of immune deficiency.

scholarly journals Embryonic Lethality and Host Immunity of RelA-Deficient Mice Are Mediated by Both Apoptosis and Necroptosis

2017 ◽  
Vol 200 (1) ◽  
pp. 271-285 ◽  
Author(s):  
Chengxian Xu ◽  
Xiaoxia Wu ◽  
Xixi Zhang ◽  
Qun Xie ◽  
Cunxian Fan ◽  
...  
Keyword(s):  
Host Immunity ◽  
Embryonic Lethality ◽  
Deficient Mice

scholarly journals Cbp Deficiency Alters Csk Localization in Lipid Rafts but Does Not Affect T-Cell Development

2005 ◽  
Vol 25 (19) ◽  
pp. 8486-8495 ◽  
Author(s):  
Shengli Xu ◽  
Jianxin Huo ◽  
Joy En-Lin Tan ◽  
Kong-Peng Lam
Keyword(s):  
T Cell ◽  
Lipid Rafts ◽  
Tyrosine Kinases ◽  
Germ Line ◽  
T Cell Development ◽  
Cell Receptor ◽  
Cell Development ◽  
Calcium Flux ◽  
Functional Defect ◽  
Deficient Mice

ABSTRACT The ubiquitously expressed transmembrane adaptor Csk-binding protein (Cbp) recruits Csk to lipid rafts, where the latter exerts its negative regulatory effect on the Src family of protein tyrosine kinases. We have inactivated Cbp in the mouse germ line. In contrast to Csk gene inactivation, which leads to embryonic lethality and impaired T-cell development, Cbp-deficient mice were viable and exhibited normal T-cell development but with an increased thymocyte population. In the absence of Cbp, the amount of Csk that localizes to the lipid rafts was greatly reduced. Interestingly, this altered lipid raft localization of Csk did not lead to any detectable biochemical or functional defect in T cells. The T-cell receptor-induced intracellular calcium flux, cell proliferation, and cytokine secretion were not affected by the absence of Cbp. Peripheral T-cell tolerance to superantigen SEB was also largely intact in Cbp-deficient mice. Thus, Cbp is dispensable for T-cell development and activation.

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