Generation of transgenic dogs that conditionally express green fluorescent protein

Kim, Min Jung; Oh, Hyun Ju; Park, Jung Eun; Hong, So Gun; Jang, Goo; Kwon, Mo Sun; Koo, Bon Chul; Kim, Teoan; Kang, Sung Keun; Chang-hyun, Jeong; Ko, CheMyong; Lee, Byeong Chun

doi:10.1002/dvg.20737

Cited by 32 publications

(13 citation statements)

References 31 publications

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“…The version of the Tet-on system used in the current study uses a single vector containing both the regulator (rtTAs-M2) and the target gene (eGFP) [21]. This vector has been previously used to generate dogs in which eGFP expression could be induced [21]. This previous study and the current study indicate that this vector could be used to generate TG animals that conditionally express proteins of interest, by replacing the eGFP gene with other genes.…”

Section: Discussionmentioning

confidence: 91%

“…In similar previous studies, a double-stable Tet-on advanced inducible cell line that contains integrated copies of the regulatory vector (pTet-On advanced) and the response vector (pTRE-Tight) was used, which is time-consuming and labor intensive. The version of the Tet-on system used in the current study uses a single vector containing both the regulator (rtTAs-M2) and the target gene (eGFP) [21]. This vector has been previously used to generate dogs in which eGFP expression could be induced [21].…”

Section: Discussionmentioning

confidence: 99%

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Production of Pigs Expressing a Transgene under the Control of a Tetracycline-Inducible System

et al. 2014

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Pigs are anatomically and physiologically closer to humans than other laboratory animals. Transgenic (TG) pigs are widely used as models of human diseases. The aim of this study was to produce pigs expressing a tetracycline (Tet)-inducible transgene. The Tet-on system was first tested in infected donor cells. Porcine fetal fibroblasts were infected with a universal doxycycline-inducible vector containing the target gene enhanced green fluorescent protein (eGFP). At 1 day after treatment with 1 µg/ml doxycycline, the fluorescence intensity of these cells was increased. Somatic cell nuclear transfer (SCNT) was then performed using these donor cells. The Tet-on system was then tested in the generated porcine SCNT-TG embryos. Of 4,951 porcine SCNT-TG embryos generated, 850 were cultured in the presence of 1 µg/ml doxycycline in vitro. All of these embryos expressed eGFP and 15 embryos developed to blastocyst stage. The remaining 4,101 embryos were transferred to thirty three surrogate pigs from which thirty eight cloned TG piglets were obtained. PCR analysis showed that the transgene was inserted into the genome of each of these piglets. Two TG fibroblast cell lines were established from these TG piglets, and these cells were used as donor cells for re-cloning. The re-cloned SCNT embryos expressed the eGFP transgene under the control of doxycycline. These data show that the expression of transgenes in cloned TG pigs can be regulated by the Tet-on/off systems.

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Section: Discussionmentioning

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Production of Pigs Expressing a Transgene under the Control of a Tetracycline-Inducible System

et al. 2014

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“…Significantly higher placental weights were observed in the cloned group (135 ± 10.1 g, n = 2), which used donor cells in confluency, compared with placentas in the natural breeding group (60 ± 7.1 g, n = 4) [16], and the cloned pups were either delivered stillborn or died within a few days after birth [16], which might be the adverse effects of placental abnormalities [17]. A second study reported a transgenic cloned dog born with a malformation, but details were not described [18]. The third report described defects in the anterior abdominal wall, and increased heart and liver sizes, muscle mass and macroglossia in 12 deceased cloned dogs that died just prior to delivery or that died with dyspnea shortly after birth [19].…”

Section: Case Presentationmentioning

confidence: 99%

Reduced birth weight, cleft palate and preputial abnormalities in a cloned dog

Kim

et al. 2014

Acta Vet Scand

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The aim of the present study was to report a novel developmental abnormality in a cloned dog. A fibroblast cell line was established from an 8-year-old male German shepherd dog. In vivo matured oocytes were retrieved from a large breed dog, and the nucleus was removed from each oocyte. A donor cell was injected into an enucleated oocyte, and the oocyte-cell couplet was fused electrically. After chemical activation, the resulting embryos were transferred into a naturally estrus-synchronized recipient dog, and two cloned pups were delivered by Cesarean section 60 days later. One cloned pup (Clone 1) was healthy, but the other (Clone 2) had a birth weight of only 320 g and cleft palate, failure of preputial closure at the ventral distal part, and persistent penile frenulum. Clone 2 was raised by stomach feeding until Day 40 after birth, where palatoplasty was performed. The abnormalities in external genitalia in Clone 2 resulted in persistent penile extrusion that was surgically corrected. This complex developmental abnormality has not been reported in dogs previously.

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“…SCNT can be performed to preserve endangered species [23], produce transgenic models [1217] and propagate dogs with elite abilities [22]. For propagation of dogs with elite abilities, seven cloned drug-sniffing dogs were produced using donor cells from a dog possessing a distinct ability to concentrate and identify specific smells among countless other scents [22].…”

Section: Introductionmentioning

confidence: 99%

Learning, memory and exploratory similarities in genetically identical cloned dogs

Shin

Park

et al. 2016

J Vet Sci

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Somatic cell nuclear transfer allows generation of genetically identical animals using donor cells derived from animals with particular traits. To date, few studies have investigated whether or not these cloned dogs will show identical behavior patterns. To address this question, learning, memory and exploratory patterns were examined using six cloned dogs with identical nuclear genomes. The variance of total incorrect choice number in the Y-maze test among cloned dogs was significantly lower than that of the control dogs. There was also a significant decrease in variance in the level of exploratory activity in the open fields test compared to age-matched control dogs. These results indicate that cloned dogs show similar cognitive and exploratory patterns, suggesting that these behavioral phenotypes are related to the genotypes of the individuals.

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Generation of transgenic dogs that conditionally express green fluorescent protein

Cited by 32 publications

References 31 publications

Production of Pigs Expressing a Transgene under the Control of a Tetracycline-Inducible System

Production of Pigs Expressing a Transgene under the Control of a Tetracycline-Inducible System

Reduced birth weight, cleft palate and preputial abnormalities in a cloned dog

Learning, memory and exploratory similarities in genetically identical cloned dogs

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