Generation of Gene Edited Pigs

Navarro-Serna, Sergio; Piñeiro-Silva, Celia; Romar, Raquel; Parrington, John; Gadea, J.

doi:10.1007/978-3-031-07496-7_3

Cited by 4 publications

(4 citation statements)

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“…The creation of pigs devoid of xenoantigens has been made possible via nuclease-based genome alteration techniques combined with somatic cell nuclear transfer (SCNT). These pigs’ organs prevent hyperacute rejection (HAR) and last longer after being transplanted into non-human primates (NHPs) ( Navarro-Serna et al, 2022 ). A clinical xenotransplantation experiment using porcine skin grafts from pigs was launched in 2019 with FDA approval.…”

Section: Nerve Repair With Tissue Engineering Scaffoldsmentioning

confidence: 99%

Nerve regeneration using decellularized tissues: challenges and opportunities

Mahdian,

Tabatabai,

Abpeikar

et al. 2023

Front. Neurosci.

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In tissue engineering, the decellularization of organs and tissues as a biological scaffold plays a critical role in the repair of neurodegenerative diseases. Various protocols for cell removal can distinguish the effects of treatment ability, tissue structure, and extracellular matrix (ECM) ability. Despite considerable progress in nerve regeneration and functional recovery, the slow regeneration and recovery potential of the central nervous system (CNS) remains a challenge. The success of neural tissue engineering is primarily influenced by composition, microstructure, and mechanical properties. The primary objective of restorative techniques is to guide existing axons properly toward the distal end of the damaged nerve and the target organs. However, due to the limitations of nerve autografts, researchers are seeking alternative methods with high therapeutic efficiency and without the limitations of autograft transplantation. Decellularization scaffolds, due to their lack of immunogenicity and the preservation of essential factors in the ECM and high angiogenic ability, provide a suitable three-dimensional (3D) substrate for the adhesion and growth of axons being repaired toward the target organs. This study focuses on mentioning the types of scaffolds used in nerve regeneration, and the methods of tissue decellularization, and specifically explores the use of decellularized nerve tissues (DNT) for nerve transplantation.

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Section: Nerve Repair With Tissue Engineering Scaffoldsmentioning

confidence: 99%

Nerve regeneration using decellularized tissues: challenges and opportunities

Mahdian,

Tabatabai,

Abpeikar

et al. 2023

Front. Neurosci.

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“…1 Percentage of embryos with 1 or more mutant alleles. 2 Percentage of mutant embryos with more than 2 alleles with respect to total embryos. 3 Percentage of mutant embryos with 1 mutant allele and 1 WT allele with respect to total embryos.…”

Section: Concentration Of Lipofectamine + Rnpmentioning

confidence: 99%

“…Therefore, they are an extremely important species agriculturally. Furthermore, their physiological and anatomical similarities to humans make pigs a great model for biomedical purposes (recently revised by Navarro-Serna et al [2]). For these reasons, genetically modified pigs are produced for a variety of applications, including the investigation of human diseases [3,4], xenotransplantation research [5,6], and the improvement of animal production [7,8].…”

Section: Introductionmentioning

confidence: 99%

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Production of Genetically Modified Porcine Embryos via Lipofection of Zona-Pellucida-Intact Oocytes Using the CRISPR/Cas9 System

Piñeiro-Silva

Navarro-Serna

Belda-Pérez

et al. 2023

Animals

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The generation of genetically modified pigs has an important impact thanks its applications in basic research, biomedicine, and meat production. Cloning was the first technique used for this production, although easier and cheaper methods were developed, such as the microinjection, electroporation, or lipofection of oocytes and zygotes. In this study, we analyzed the production of genetically modified embryos via lipofection of zona-pellucida-intact oocytes using LipofectamineTM CRISPRMAXTM Cas9 in comparison with the electroporation method. Two factors were evaluated: (i) the increment in the concentration of the lipofectamine–ribonucleoprotein complexes (LRNPC) (5% vs. 10%) and (ii) the concentration of ribonucleoprotein within the complexes (1xRNP vs. 2xRNP). We found that the increment in the concentration of the LRNPC had a detrimental effect on embryo development and a subsequent effect on the number of mutant embryos. The 5% group had a similar mutant blastocyst rate to the electroporation method (5.52% and 6.38%, respectively, p > 0.05). The increment in the concentration of the ribonucleoprotein inside the complexes had no effect on the blastocyst rate and mutation rate, with the mutant blastocyst rate being similar in both the 1xRNP and 2xRNP lipofection groups and the electroporation group (1.75%, 3.60%, and 3.57%, respectively, p > 0.05). Here, we showed that it is possible to produce knock-out embryos via lipofection of zona-pellucida-intact porcine oocytes with similar efficiencies as with electroporation, although more optimization is needed, mainly in terms of the use of more efficient vesicles for encapsulation with different compositions.

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