Generation of Transgenic Pigs Overexpressing PID1 Gene
Mediated by Magnetic Nanoparticles and Sperm

Fang, Guo-feng; Chen, Wei; Wang, Shoudong; Wang, Yandong; Li, Chuanhao; Zhu, Haibo; Wang, Hui; Zeng, Yongqing

doi:10.3923/ajava.2017.161.168

Cited by 3 publications

(3 citation statements)

References 25 publications

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“…PID1‐transgenic pigs were genetated by sperm and magnetic nanoparticles‐mediated method as described . Seven PID1‐transgenic pigs were identified by polymerase chain reaction (PCR), Southern blot, quantitative real‐time PCR, and western‐blot …”

Section: Methodsmentioning

confidence: 99%

Overexpression of PID1 reduces high density lipoprotein level and functionality in swine

Zhu

et al. 2019

IUBMB Life

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Phosphotyrosine interaction domain 1 (PID1), a protein with a phosphotyrosinebinding (PTB) domain, interacts with the lipoprotein receptor-related protein 1 (LRP1) to reduce the insulin sensitivity of adipocyte. Considering the role of LRP1 in lipid metabolism, we investigated the effect of PID1 on the content and biological activities of serum lipoproteins in pigs. PID1-transgenic pigs were genetated by sperm and magnetic nanoparticles-mediated method. The levels of PID1 in PID1-transgenic pig's liver were higher than that in the wild-type pig's liver. We found that serum levels of highdensity lipoprotein cholesterol (HDL-C) and apolipoprotein A-I (apoA-I) were significantly reduced in PID1-transgenic pigs. On the other hand, PID1-transgenic pigs displayed increased non-HDL-C levels. Serum levels of total cholesterol and triglycerides were comparable between the PID1-transgenic and the wild-type pigs. Further, the HDL isolated from PID1-transgenic pigs showed a significant reduction in cholesterol efflux ability. In addition, serum superoxide dismutase activity of PID1-transgenic pigs was also obviously lowered compared with that of wild type pigs. In conclusion, these results suggest that PID1 might be able to adjust HDL-C levels in serum and HDL cholesterol efflux ability.Abbreviations: HDL-C, high-density lipoprotein cholesterol; LCAT, Lecithin-cholesterol acyltransferase; LDL-C, low density lipoprotein cholesterol; LRP1, lipoprotein receptor-related protein 1; PID1, Phosphotyrosine interaction domain 1; SOD, serum superoxide dismutase; TC, total cholesterol; TG, triglycerides; VLDL, very low-density lipoprotein.

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Section: Methodsmentioning

confidence: 99%

Overexpression of PID1 reduces high density lipoprotein level and functionality in swine

Zhu

et al. 2019

IUBMB Life

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“…The spermatozoon internalizes DNA more easily than most cells. A recent study (Fang et al ., 2017) used approximately 500 μl of 800 mg/ml plasmid solution and an equal volume (500 μl) of polyMAG-1000 for transfection of porcine spermatozoa, however transfection rates and sperm viability were not measured.…”

Section: Discussionmentioning

confidence: 99%

“…PLF plus PEI has been evaluated previously in some cell types (Dang et al ., 2011; Hsu & Uludag, 2012). For pig spermatozoa, PEI-coated magnetic nanoparticles (MNP-PEI) alone were used (Fang et al ., 2017), however PEI polyplex itself has not been tested on sperm from animal species. Using EP and PLF as the transfection method, various success rates have been observed in different cell types including spermatozoa from many species but not from swine.…”

Section: Introductionmentioning

confidence: 99%

Comparison between electroporation and polyfection in pig sperm: efficiency and cell viability implications

Silva

Souza

Pandolfi

et al. 2018

Zygote

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SummaryThe aim of this study was to optimize protocols for electroporation (EP) and polyfection (PLF) using polyethyleneimine (PEI) for pig sperm transfection and to determine which method was the most efficient. For EP standardization, different voltages, amounts and times of electric pulses were tested using propidium iodide (PI) as reporter. For PLF standardization, different concentrations of fluorescein isothiocyanate (FITC)-labelled PEI (PEI/FITC) were incubated with sperm for different periods of time. Flow cytometry was performed to evaluate the best protocol in terms of cell viability, including cytoplasmic membrane, acrosome, chromatin integrities and mitochondrial potential using the FITC probe, PI, acridine orange (AO) and JC1. Transfections with the plasmid pmhyGENIE-5 were carried out under optimum conditions for each procedure (EP: 500 volts, 500 μs and two pulses; PLF: PEI 0.5 mg/ml and incubation time 10 min). Transfection efficacy was assessed by fluorescence in situ hybridization (FISH). A lower transfection rate was observed for sperm in the control group (17.8%) compared with EP (36.7%), with PLF (76.8%) being the most efficient. These results suggest that the EP and PEI could be an efficient and low cost transfection method for swine sperm. Notably, treated cells showed higher plasmatic the membrane damage (PMD) and/or acrosome damage (AD) indexes, therefore the combination of this procedure with biotechniques that facilitate fecundation (i.e. in vitro fertilization or intracytoplasmic sperm injection) or even inclusion of antioxidant or anti-apoptotic drugs to improve spermatozoa viability would be important.

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Advances in transgene delivery for the generation of transgenic livestock

Pramod

Mitra²

2023

Nanobiotechnology for the Livestock Industry

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Generation of Transgenic Pigs Overexpressing PID1 Gene Mediated by Magnetic Nanoparticles and Sperm

Cited by 3 publications

References 25 publications

Overexpression of PID1 reduces high density lipoprotein level and functionality in swine

Overexpression of PID1 reduces high density lipoprotein level and functionality in swine

Comparison between electroporation and polyfection in pig sperm: efficiency and cell viability implications

Advances in transgene delivery for the generation of transgenic livestock

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