Refinements in embryo manipulation applied to CRISPR technology in livestock

Santos-Neto, P.C. dos; Cuadro, F.; Souza-Neves, Marcela; Crispo, Martina; Menchaca, A.

doi:10.1016/j.theriogenology.2023.05.028

Cited by 2 publications

(1 citation statement)

References 24 publications

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“…This has facilitated the generation of additional genetically engineered rat models for skeletal diseases [ 140 , 141 ]. In addition, while large animal models still require more resources and time to establish and maintain, they are more predictive regarding pre-clinical therapeutic modeling [ 142 , 143 , 144 , 145 , 146 , 147 ]. CRISPR/Cas9 and other recent approaches will undoubtedly accelerate the development of larger animal models for skeletal diseases.…”

Section: Discussionmentioning

confidence: 99%

The Past, Present, and Future of Genetically Engineered Mouse Models for Skeletal Biology

Michalski,

Williams

2023

Biomolecules

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The ability to create genetically engineered mouse models (GEMMs) has exponentially increased our understanding of many areas of biology. Musculoskeletal biology is no exception. In this review, we will first discuss the historical development of GEMMs and how these developments have influenced musculoskeletal disease research. This review will also update our 2008 review that appeared in BONEKey, a journal that is no longer readily available online. We will first review the historical development of GEMMs in general, followed by a particular emphasis on the ability to perform tissue-specific (conditional) knockouts focusing on musculoskeletal tissues. We will then discuss how the development of CRISPR/Cas-based technologies during the last decade has revolutionized the generation of GEMMs.

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Section: Discussionmentioning

confidence: 99%

The Past, Present, and Future of Genetically Engineered Mouse Models for Skeletal Biology

Michalski,

Williams

2023

Biomolecules

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Microsurgical separation of deconserved embryos

Babenkov,

Ubushieva,

Chimidova

et al. 2024

E3S Web Conf.

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At the current level of development of cell engineering methods in cattle breeding, the accelerated production of monozygotic twins is acquiring important scientific and economic significance. This will make it possible to determine the effectiveness of breeding methods in assessing genetic progress in a population over a number of generations. The method of embryonic cloning by microsurgical separation of embryos is close to solving this problem. Of particular interest is the long-term preservation of demi-embryos to obtain genetic analogues. But the possibilities of preserving demi-embryos at low temperatures are limited, since existing methods are multi-stage and extremely difficult to implement in practice. The search for applied methods of microsurgery in combination with long-term preservation of demi-embryos is an urgent task. Early studies have reported micromanipulation of frozen-thawed embryos. In 1984, in experiments by S. Willadsen & R. Godke, after microsurgery of deconserved embryos, the survival rate was 54.2%. Data on survival rate are not provided. No other data was found, and therefore we conducted studies to determine the influence of the processes of cryo- and deconservation, and dissection of embryos on the survival and ability of demi-embryos to develop totipotently after transplantation. The research was carried out in 2022 at the Regional Research and Production Center for the Reproduction of Farm Animals of Kalmyk State University named after B.B. Gorodovikov. The efficiency of dissection of intact embryos was 86.1% - after separation of 61 embryos, 105 suitable demi-embryos were obtained out of 122 possible, which was 5.7% higher than the same figure for dividing deconserved embryos - 80.4% (37 suitable demi-embryos out of 46 possible after separation 23 embryos). The survival rate of demi-embryos as the ability to nidate in the recipient's uterus in intact and depreserved demi-embryos differed within acceptable limits - 52.2% when transplanting 1 intact demi-embryo (12 pregnant out of 23 recipients) and 65.9% when transplanting 2 demi-embryos (27 pregnant from 41). Similar indicators for transplantation of deconserved demi-embryos were respectively: 42.8 (3 pregnant recipients out of 7 transplanted) and 60.0% (9 pregnant recipients out of 15).

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Refinements in embryo manipulation applied to CRISPR technology in livestock

Cited by 2 publications

References 24 publications

The Past, Present, and Future of Genetically Engineered Mouse Models for Skeletal Biology

The Past, Present, and Future of Genetically Engineered Mouse Models for Skeletal Biology

Microsurgical separation of deconserved embryos

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