Emergency prevention of extinction of a transgenic allele in a less-fertile transgenic mouse line by crossing with an inbred or outbred mouse strain coupled with assisted reproductive technologies

2007 ◽  
Vol 19 (8) ◽  
pp. 984 ◽  
Author(s):  
Anna Mayer ◽  
Diana Bulian ◽  
Hagen Scherb ◽  
Martin Hrabé de Angelis ◽  
Jörg Schmidt ◽  
...  
Keyword(s):  
Transgenic Mouse ◽  
Fluorescent Protein ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Low Fertility ◽  
In Vitro Fertilisation ◽  
Mouse Line ◽  
Transgenic Mouse Line ◽  
Green Fluorescent

Certain transgenic mouse lines are difficult to breed or archive and, consequently, their transgenes become lost. We examined a C57BL/6 mouse line (B6-tg), transgenic for green fluorescent protein (GFP) with low fertility, and its crosses with the more prolific inbred C3HeB/FeJ (C3) and outbred Swiss (SW) strains in order to assess the possibility of emergency prevention of extinction of a transgenic allele by using assisted reproductive technologies (ART). Out-crossing was performed by natural mating or in vitro fertilisation (IVF) with heterozygous mice. Most of the crossing combinations resulted in improved archiving and rederivation efficiencies of the transgenic allele. Natural crossing increased both mean litter size by two to three pups and the superovulatory rate from 69% for B6-tg to 70–90% for females from the out-crosses. Each plug-positive B6-tg female yielded an average of 4.6 two-cell embryos, whereas females from the out-crosses produced three- to fivefold that amount. After thawing, 13% of B6-tg embryos and 6–12% of out-crossed embryos developed into transgenic pups after transfer into recipients. After IVF with cryopreserved spermatozoa, cleavage rates were 4% for B6-tg, 22–37% for B6-tg oocytes out-crossed with C3 and SW spermatozoa, 9–49% for gametes from out-crossed mice and 28–44% for back-crosses with B6 oocytes. Transgenic pups were not derived from IVF with B6-tg gametes when either fresh or cryopreserved spermatozoa were used. Rederivation efficiencies were 7% and 4% from out-crosses of B6-tg oocytes with C3 and SW spermatozoa, respectively, 6–22% for gametes from out-crossed mice and 4–10% for the back-crosses. Although out-crossing changes the original genetic background, the strategy of crossing coupled with ART prevents the extinction of an allele of interest, especially where archiving and rederivation of the transgenic line fail.

scholarly journals A New Transgenic Mouse Line for Imaging Mitochondrial Calcium Signals

Function ◽  
2021 ◽  
Vol 2 (3) ◽  
Author(s):  
Nelly Redolfi ◽  
Elisa Greotti ◽  
Giulia Zanetti ◽  
Tino Hochepied ◽  
Cristina Fasolato ◽  
...  
Keyword(s):  
Transgenic Mouse ◽  
Fluorescent Protein ◽  
Ex Vivo ◽  
Brain Slices ◽  
Resonance Energy ◽  
Mouse Line ◽  
Isolated Cells ◽  
Genomic Locus ◽  
Transgenic Mouse Line

AbstractMitochondria play a key role in cellular calcium (Ca2+) homeostasis. Dysfunction in the organelle Ca2+ handling appears to be involved in several pathological conditions, ranging from neurodegenerative diseases, cardiac failure and malignant transformation. In the past years, several targeted green fluorescent protein (GFP)-based genetically encoded Ca2+ indicators (GECIs) have been developed to study Ca2+ dynamics inside mitochondria of living cells. Surprisingly, while there is a number of transgenic mice expressing different types of cytosolic GECIs, few examples are available expressing mitochondria-localized GECIs, and none of them exhibits adequate spatial resolution. Here we report the generation and characterization of a transgenic mouse line (hereafter called mt-Cam) for the controlled expression of a mitochondria-targeted, Förster resonance energy transfer (FRET)-based Cameleon, 4mtD3cpv. To achieve this goal, we engineered the mouse ROSA26 genomic locus by inserting the optimized sequence of 4mtD3cpv, preceded by a loxP-STOP-loxP sequence. The probe can be readily expressed in a tissue-specific manner upon Cre recombinase-mediated excision, obtainable with a single cross. Upon ubiquitous Cre expression, the Cameleon is specifically localized in the mitochondrial matrix of cells in all the organs and tissues analyzed, from embryos to aged animals. Ca2+ imaging experiments performed in vitro and ex vivo in brain slices confirmed the functionality of the probe in isolated cells and live tissues. This new transgenic mouse line allows the study of mitochondrial Ca2+ dynamics in different tissues with no invasive intervention (such as viral infection or electroporation), potentially allowing simple calibration of the fluorescent signals in terms of mitochondrial Ca2+ concentration ([Ca2+]).

scholarly journals A Tendon-Specific Double Reporter Transgenic Mouse Enables Tracking Cell Lineage and Functions Alteration In Vitro and In Vivo

2021 ◽  
Vol 22 (20) ◽  
pp. 11189
Author(s):  
Rui Chen ◽  
Xunlei Zhou ◽  
Thomas Skutella
Keyword(s):  
Transgenic Mouse ◽  
Fluorescent Protein ◽  
Cell Lineage ◽  
Mouse Line ◽  
Reporter System ◽  
Specific Expression ◽  
Transgenic Mouse Line ◽  
Blue Fluorescent Protein

We generated and characterized a transgenic mouse line with the tendon-specific expression of a double fluorescent reporter system, which will fulfill an unmet need for animal models to support real-time monitoring cell behaviors during tendon development, growth, and repair in vitro and in vivo. The mScarlet red fluorescent protein is driven by the Scleraxis (Scx) promoter to report the cell lineage alteration. The blue fluorescent protein reporter is expressed under the control of the 3.6kb Collagen Type I Alpha 1 Chain (Col1a1) proximal promoter. In this promoter, the existence of two promoter regions named tendon-specific cis-acting elements (TSE1, TSE2) ensure the specific expression of blue fluorescent protein (BFP) in tendon tissue. Collagen I is a crucial marker for tendon regeneration that is a major component of healthy tendons. Thus, the alteration of function during tendon repair can be estimated by BFP expression. After mechanical stimulation, the expression of mScarlet and BFP increased in adipose-derived mesenchymal stem cells (ADMSCs) from our transgenic mouse line, and there was a rising trend on tendon key markers. These results suggest that our tendon-specific double reporter system is a novel model used to study cell re-differentiation and extracellular matrix alteration in vitro and in vivo.

scholarly journals May I have your uterus? The contribution of considering complexities preceding live uterus transplantation

2021 ◽  
pp. medhum-2020-011864
Author(s):  
Lisa Guntram
Keyword(s):  
Medical Humanities ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
In Vitro Fertilisation ◽  
Medical Innovation ◽  
Uterus Transplantation ◽  
Ethical Debate ◽  
Care And Support ◽  
Different Dimensions

Uterus transplantation combined with in vitro fertilisation (IVF) (henceforth called UTx-IVF) as a treatment for infertility caused by an absence or malfunction of the uterus is advancing. About 50 transplantations have been conducted worldwide and at least 14 children have been born—9 of them by women taking part in a Swedish research project on UTx-IVF. The Swedish research protocol initially stated that the potential recipient must ‘have her own donor’ who is preferably related to the recipient. But what does it mean to ask someone for a uterus? What challenges does this question instigate? And what norms may it enact? In this article, I explore how 10 women—who have considered, and sometimes pursued, UTx-IVF—describe their experiences of searching for a donor. I aim to show how an analysis of such accounts can help us unpack some of the specific relational and gendered dimensions of UTx-IVF and by doing so enrich discussions of risks, benefits, care and support in UTx-IVF. Drawing on research in social sciences and medical humanities that has demonstrated how assisted reproductive technologies and organ donation can provoke social and familial conundrums, with respect to such topics as embodiment and identity, I present three patterns that describe different dimensions of the interviewees’ quest for a uterus donor. I discuss the negotiations that took place, how expectations unfolded and how entanglements were managed as the interviewees considered asking someone for a donation. Such an examination, I suggest, contributes to make care and support more attuned to the experiences and entanglements that UTx-IVF entails for those pursuing it. This will become increasingly important if (or when) UTx-IVF becomes part of general healthcare. To conclude, I problematise responsibilities and relational challenges in medical innovation, and in this way provide insights into how the ethical debate over UTx-IVF can broaden its scope.

scholarly journals A Transgenic Mouse Line Expressing the Red Fluorescent Protein tdTomato in GABAergic Neurons

PLoS ONE ◽  
2015 ◽  
Vol 10 (6) ◽  
pp. e0129934 ◽  
Author(s):  
Stefanie Besser ◽  
Marit Sicker ◽  
Grit Marx ◽  
Ulrike Winkler ◽  
Volker Eulenburg ◽  
...  
Keyword(s):  
Transgenic Mouse ◽  
Fluorescent Protein ◽  
Gabaergic Neurons ◽  
Mouse Line ◽  
Red Fluorescent Protein ◽  
Transgenic Mouse Line

Rare occurrence of Tetralogy of Fallot in dizygotic twins conceived via in vitro fertilisation

2019 ◽  
Vol 29 (12) ◽  
pp. 1541-1542
Author(s):  
Jason W. Greenberg ◽  
Chetana Reddy ◽  
Charles B. Huddleston
Keyword(s):  
Tetralogy Of Fallot ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
In Vitro Fertilisation ◽  
Rare Occurrence ◽  
Dizygotic Twins

AbstractAn increased incidence of CHD has been noted in twin gestations and in infants conceived using assisted reproductive technologies. However, CHD in these populations remains understudied and the mechanisms underlying these phenomena remain unclear. We present the case of twins conceived via in vitro fertilisation both with Tetralogy of Fallot and additional cardiac and extracardiac malformations.

Is it ethical to provide IVF add-ons when there is no evidence of a benefit if the patient requests it?

2019 ◽  
Vol 45 (5) ◽  
pp. 346-350 ◽  
Author(s):  
Mila Stefanova Zemyarska
Keyword(s):  
Assisted Reproductive Technologies ◽  
Cost Effective ◽  
Reproductive Technologies ◽  
Conclusive Evidence ◽  
Diagnostic Tools ◽  
In Vitro Fertilisation ◽  
Unborn Child ◽  
Effective Adjunct ◽  
Psychological Benefit

In vitro fertilisation (IVF) ‘add-ons’ are therapeutic or diagnostic tools developed in an endeavour to improve the success rate of infertility treatment. However, there is no conclusive evidence that these interventions are a beneficial or effective adjunct of assisted reproductive technologies. Additionally, IVF add-ons are often implemented in clinical practice before their safety can be thoroughly ascertained. Yet, patients continue to request and pay large sums for such additional IVF tools. Hence, this essay set out to examine if it is ethical to provide IVF add-ons when there is no evidence of a benefit if the patient requests it. In order to determine what is ethical—namely, morally good and righteous, the question was considered in relation to three key values of medical ethics—autonomy, beneficence and non-maleficence. It was determined that providing IVF add-ons might be morally acceptable in specific circumstances, if true informed consent can be given, there is a potential of cost-effective physiological or psychological benefit and the risk of harm is minimal, particularly with regard to the unborn child.

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