Minimum volume Spatula MVD vitrification method improves embryo survival compared to traditional slow freezing, both for in vivo and in vitro produced mice embryos

Meikle, María Noel; Schlapp, Geraldine; Menchaca, A.; Crispo, Martina

doi:10.1016/j.cryobiol.2018.07.005

Cited by 13 publications

(13 citation statements)

References 23 publications

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“…The outcomes on 1207 embryos microinjected with CRISPR that were transferred in different developmental stages, associated or not to vitrification, are presented in Table 1. Vitrification by minimum volume methods (Crytop and Spatula MVD) are used as a routine in our laboratory with acceptable outcomes, both in livestock and murine embryos after microinjection for genome modification [32,41]. This practice simplifies embryo transfer programs to produce genome-edited animals, mainly in large-scale projects involving livestock.…”

Section: Embryo Manipulationmentioning

confidence: 99%

CRISPR in livestock: From editing to printing

et al. 2020

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a b s t r a c tPrecise genome editing of large animals applied to livestock and biomedicine is nowadays possible since the CRISPR revolution. This review summarizes the latest advances and the main technical issues that determine the success of this technology. The pathway from editing to printing, from engineering the genome to achieving the desired animals, does not always imply an easy, fast and safe journey. When applied in large animals, CRISPR involves time-and cost-consuming projects, and it is mandatory not only to choose the best approach for genome editing, but also for embryo production, zygote microinjection or electroporation, cryopreservation and embryo transfer. The main technical refinements and most frequent questions to improve this disruptive biotechnology in large animals are presented. In addition, we discuss some CRISPR applications to enhance livestock production in the context of a growing global demand of food, in terms of increasing efficiency, reducing the impact of farming on the environment, enhancing pest control, animal welfare and health. The challenge is no longer technical. Controversies and consensus, opportunities and threats, benefits and risks, ethics and science should be reconsidered to enter into the CRISPR era.

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Section: Embryo Manipulationmentioning

confidence: 99%

CRISPR in livestock: From editing to printing

et al. 2020

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“…Further dissecting the biological role of CCDC28B will likely shed important insight to understand not only BBS but also ASD. pseudo-pregnant females (25 embryos/female in average), following surgical procedures established in our animal facility [68]. For surgery, recipient females were anesthetized with a mixture of ketamine (100 mg/kg, Pharmaservice, Ripoll Vet, Montevideo, Uruguay) and xylazine (10 mg/kg, Seton 2%; Calier, Montevideo, Uruguay).…”

Section: Discussionmentioning

confidence: 99%

Generation and characterization ofCcdc28bmutant mice links the Bardet-Biedl associated gene with social behavioral phenotypes

Fabregat

Niño

Pose

et al. 2021

Preprint

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CCDC28B (coiled-coil domain-containing protein 28B) was identified as a modifier in the ciliopathy Bardet-Biedl syndrome (BBS). Our previous work in cells and zebrafish showed that CCDC28B plays a role regulating cilia length in a mechanism that is not completely understood. Here we report the generation of a Ccdc28b mutant mouse using CRISPR/Cas9 (Ccdc28b mut). After confirming the depletion of Ccdc28b ;we performed a phenotypic characterization showing that Ccdc28b mut animals present a mild phenotype: i) do not present clear structural cilia affectation, although we did observe mild defects in cilia density and cilia length in some tissues, ii) reproduce normally, and iii) do not develop retinal degeneration or obesity, two hallmark features of reported BBS murine models. In contrast, Ccdc28b mut mice did show clear social interaction defects as well as stereotypical behaviors suggestive of autism spectrum disorder (ASD). This finding is indeed relevant regarding CCDC28B as a modifier of BBS since behavioral phenotypes have been documented in BBS. Importantly however, our data suggests a possible causal link between CCDC28B and ASD-like phenotypes that exceeds the context of BBS: filtering for rare deleterious variants, we found CCDC28B mutations in eight probands from the Simmons Simplex Collection cohort. Furthermore, a phenotypic analysis showed that CCDC28B mutation carriers present lower BMI and mild communication defects compared to a randomly selected sample of SSC probands. Thus, our results suggest that mutations in CCDC28B lead to mild autism-like features in mice and humans. Overall, this work reports a novel mouse model that will be key to continue evaluating genetic interactions in BBS, deciphering the contribution of CCDC28B to modulate the presentation of BBS phenotypes. In addition, our data underscores a novel link between CCDC28B and ASD-like phenotypes, providing a novel opportunity to further our understanding of the genetic, cellular, and molecular basis of ASD.

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“…ET was conducted in a randomly blocked design for each treatment (EE and control group), regarding embryo stage (956 zygotes and 465 morulae) and the operator (two trained technicians). Embryo transfer, surgical care and pain relief with 1 mg/kg Tolfenamic acid was performed as previously described [ 19 , 20 ]. Embryos were loaded into a pulled glass pipette and placed through the infundibulum into both oviducts, delivering in average 25–30 zygotes or 17–20 morulae per female.…”

Section: Methodsmentioning

confidence: 99%

Long-Term Effect of Environmental Enrichment on Reproductive Performance of Swiss Webster Mice and Their Female Offspring

Meikle

Arevalo

Schlapp

et al. 2020

Animals

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The aim of this study was to evaluate the effect of an environmental enrichment (EE) plan on the reproductive performance of Swiss Webster mice and their female offspring used as recipients for embryo transfer. A total of 54 breeder mice and 60 F1 females, used as foster mothers, were allocated in two experimental groups to receive or not receive EE for physical well-being. Reproductive outcomes of the Swiss trios such as birth rate and pup number, litter size, pups’ weight at weaning, interlitter interval and time to first litter were analyzed. Environmental enrichment significantly increased pups weight from breeding trios compared to the control group (14.4 ± 0.1 vs. 13.8g ± 0.1, EE vs. control, respectively; p < 0.01). Other parameters did not differ between both groups. Reproductive parameters of female offspring used as recipients for embryo transfer did not differ among groups subjected or not to EE. These data demonstrate that the EE protocol applied in Swiss Webster breeder mice positively enhanced pups weight, and did not interfere with other reproductive outcomes. In conclusion, this study supports the implementation of EE plans usually applied for animal welfare in mouse facilities with slight improvement in reproductive performance.

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Minimum volume Spatula MVD vitrification method improves embryo survival compared to traditional slow freezing, both for in vivo and in vitro produced mice embryos

Cited by 13 publications

References 23 publications

CRISPR in livestock: From editing to printing

CRISPR in livestock: From editing to printing

Generation and characterization ofCcdc28bmutant mice links the Bardet-Biedl associated gene with social behavioral phenotypes

Long-Term Effect of Environmental Enrichment on Reproductive Performance of Swiss Webster Mice and Their Female Offspring

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