Human lactoferrin efficiently targeted into caprine beta-lactoglobulin locus with transcription activator-like effector nucleases

Yuan, Yawei; Song, Shaozheng; Zhu, Meng-Ming; He, Zheng-yi; Lü, Rui; Zhang, Ting; Mi, Fei; Wang, Jinyu; Cheng, Yiwei

doi:10.5713/ajas.16.0697

Cited by 11 publications

(13 citation statements)

References 28 publications

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“…This initial report presented the potential of the TALEN endonucleases for the introduction of desirable allelic introgressions into the genomes of farm animals. TALENs have also been applied in goats to target the BLG gene (Cui et al, 2015; Ge et al, 2016; Zhu et al, 2016b; Yuan et al, 2017). Furthermore, gene-edited goats that carry both BLG knockout and enriched expression of human lactoferrin (hLF) (Cui et al, 2015) or human α-lactalbumin (hLA) (Zhu et al, 2016b) have been generated using SCNT.…”

Section: Overview Of Genetic Modification and Relevant Biotechnologicmentioning

confidence: 99%

Sheep and Goat Genome Engineering: From Random Transgenesis to the CRISPR Era

et al. 2019

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Sheep and goats are valuable livestock species that have been raised for their production of meat, milk, fiber, and other by-products. Due to their suitable size, short gestation period, and abundant secretion of milk, sheep and goats have become important model animals in agricultural, pharmaceutical, and biomedical research. Genome engineering has been widely applied to sheep and goat research. Pronuclear injection and somatic cell nuclear transfer represent the two primary procedures for the generation of genetically modified sheep and goats. Further assisted tools have emerged to enhance the efficiency of genetic modification and to simplify the generation of genetically modified founders. These tools include sperm-mediated gene transfer, viral vectors, RNA interference, recombinases, transposons, and endonucleases. Of these tools, the four classes of site-specific endonucleases (meganucleases, ZFNs, TALENs, and CRISPRs) have attracted wide attention due to their DNA double-strand break-inducing role, which enable desired DNA modifications based on the stimulation of native cellular DNA repair mechanisms. Currently, CRISPR systems dominate the field of genome editing. Gene-edited sheep and goats, generated using these tools, provide valuable models for investigations on gene functions, improving animal breeding, producing pharmaceuticals in milk, improving animal disease resistance, recapitulating human diseases, and providing hosts for the growth of human organs. In addition, more promising derivative tools of CRISPR systems have emerged such as base editors which enable the induction of single-base alterations without any requirements for homology-directed repair or DNA donor. These precise editors are helpful for revealing desirable phenotypes and correcting genetic diseases controlled by single bases. This review highlights the advances of genome engineering in sheep and goats over the past four decades with particular emphasis on the application of CRISPR/Cas9 systems.

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Section: Overview Of Genetic Modification and Relevant Biotechnologicmentioning

confidence: 99%

Sheep and Goat Genome Engineering: From Random Transgenesis to the CRISPR Era

et al. 2019

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“…CFFCs were isolated from 70-day-old fetuses that were recovered surgically from a Boer goat obtained from the Yangzhou University farm, as previously described [ 27 ]. Briefly, pregnant goats were anesthetized using an intramuscular injection of serazine hydrochloride (0.02 mL/kg body weight), and fetuses were collected.…”

Section: Methodsmentioning

confidence: 99%

Graphene Oxide–Silver Nanoparticle Nanocomposites Induce Oxidative Stress and Aberrant Methylation in Caprine Fetal Fibroblast Cells

Yuan

Cai

Wang

et al. 2021

Cells

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Graphene oxide–silver nanoparticle (GO-AgNPs) nanocomposites have drawn much attention for their potential in biomedical uses. However, the potential toxicity of GO-AgNPs in animals and humans remains unknown, particularly in the developing fetus. Here, we reported the GO-AgNP-mediated cytotoxicity and epigenetic alteration status in caprine fetal fibroblast cells (CFFCs). In brief, the proliferation and apoptosis rate of GO-AgNP-treated CFFCs (4 and 8 µg/mL of GO-AgNPs) were measured using the cell-counting kit (CCK-8) assay and the annexin V/propidium iodide (PI) assay, respectively. In addition, the oxidative stress induced by GO-AgNPs and detailed mechanisms were studied by evaluating the generation of reactive oxygen species (ROS), superoxide dismutase (SOD), lactate dehydrogenase (LDH), malondialdehyde (MDA), and caspase-3 and abnormal methylation. The expression of pro- and anti-apoptotic genes and DNA methyltransferases was measured using reverse transcription followed by RT-qPCR. Our data indicated that GO-AgNPs cause cytotoxicity in a dose-dependent manner. GO-AgNPs induced significant cytotoxicity by the loss of cell viability, production of ROS, increasing leakage of LDH and level of MDA, increasing expression of pro-apoptotic genes, and decreasing expression of anti-apoptotic genes. GO-AgNPs incited DNA hypomethylation and the decreased expression of DNMT3A. Taken together, this study showed that GO-AgNPs increase the generation of ROS and cause apoptosis and DNA hypomethylation in CFFCs. Therefore, the potential applications of GO-AgNPs in biomedicine should be re-evaluated.

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“…Caprine fetal broblast cells were isolated from 70-day old fetuses that were recovered surgically from a Boer goat obtained from Yangzhou University farm as previously described [57]. Brie y, the pregnant goats were anaesthetized using intramuscular injected with serazine hydrochloride (0.02 mL/kg body weight) and fetuses were collected.…”

Section: Cell Culturementioning

confidence: 99%

Graphene oxide-silver nanocomposites induce oxidative stress and apoptosis in caprine fetal fibroblast cells

Yuan¹,

Joo²,

Wang³

et al. 2020

Preprint

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Background: Graphene oxide (GO) has drawn much attention as excellent platform to which silver nanoparticles (AgNPs) can be anchored for the production of biomedical nanocomposites. Yet, the potential toxicity of reduced graphene oxide- silver nanoparticles (GO-AgNPs) nanocomposites to animal and human is complex to evaluate and remains largely unknown.Results: Our data indicated that GO-AgNPs caused cytotoxicity in dose-dependent manner. GO-AgNPs induced significant cytotoxicity by the loss of cell viability, over-production of reactive oxygen species (ROS), increased leakage of lactate dehydrogenase (LDH) and level of malondialdehyde (MDA), increased expression of pro-apoptotic genes and decreased expression of anti-apoptotic genes.Conclusions: This study demonstrated that GO-AgNPs potentially induced oxidative stress, which resulted in toxicity and cell apoptosis in caprine fetal fibroblast cell due to an increased generation of ROS. For antibacterial applications of GO-AgNPs nanocomposite in animal, the toxical effect must be further evaluated.

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Human lactoferrin efficiently targeted into caprine beta-lactoglobulin locus with transcription activator-like effector nucleases

Cited by 11 publications

References 28 publications

Sheep and Goat Genome Engineering: From Random Transgenesis to the CRISPR Era

Sheep and Goat Genome Engineering: From Random Transgenesis to the CRISPR Era

Graphene Oxide–Silver Nanoparticle Nanocomposites Induce Oxidative Stress and Aberrant Methylation in Caprine Fetal Fibroblast Cells

Graphene oxide-silver nanocomposites induce oxidative stress and apoptosis in caprine fetal fibroblast cells

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