CMV enhancer–promoter is preferentially active in exocrine cells in vivo

Vasey, Douglas B.; Lillico, Simon; Sang, Helen; King, Tim; Whitelaw, Bruce

doi:10.1007/s11248-008-9235-y

Cited by 13 publications

(18 citation statements)

References 20 publications

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“…Presence of ZsGreen1 in the skin of neonatal transgenic piglets was detected visually (Figure 1A) and via immunoblot ( Figure 1B), indicative of CMV promoter activity. Even without the UV light, transgenic animals were identifiable via the yellow hue of their skin (data not shown), which has been previously noted in GFP pigs [26]. The genotype of the animals was confirmed by 185 conventional PCR ( Figure 1D); a schematic depicting the genotyping method is available in Figure 1C.…”

Section: Identification Of Hemizygous and Homozygous Cmv-zsgreen1 Trasupporting

confidence: 70%

“…The CMV promoter has been utilized to drive transgene expression in numerous vertebrate animal models including: sheep [15][16][17], fish [14,18], mice [19][20][21][22], rats [23,24], chickens [25] and pigs [4,[26][27][28]. In the pig, the CMV promoter was used to drive expression of green fluorescent protein (GFP) [27].…”

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confidence: 99%

“…As anticipated, expression of the transgene was detected in a wide range of organs, including those derived from each of the 3 germ layers: skin (ectoderm), pancreas 65 (endoderm), and kidney (mesoderm). Despite apparent universal transgene expression [27], the CMV promoter was preferentially active in exocrine vs. non-exocrine cells of the pig [26]. Given the growing interest in the development of transgenic swine models, the objective of this study was to quantify activity of the CMV promoter in a wide range of porcine tissues.…”

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confidence: 99%

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A transgenic pig model expressing a ZsGreen1 reporter across an extensive array of tissues

Desaulniers

Cederberg

Carreiro

et al. 2018

Preprint

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The advent of genetically engineered pig production has revealed a wide array of opportunities to enhance both biomedical and agricultural industries. One powerful method to develop these models is transgenesis; however, selection of a suitable promoter to drive transgene expression is critical. The cytomegalovirus (CMV) promoter is the most commonly used viral promoter as it robustly drives transgene expression in a ubiquitous nature. However, recent reports suggest that the level of CMV promoter activity is tissue-dependent in the pig. Therefore, the objective of this study was to quantify the activity of the CMV promoter in a wide range of porcine tissues. Swine harboring a CMV-ZsGreen1 transgene were utilized for this study. Thirty five tissue samples were collected from neonatal hemizygous (n = 3) and homozygous (n = 3) transgenic piglets and analyzed for ZsGreen1 abundance via immunoblot. ZsGreen1 was detected in all tissues examined; however, quantification revealed that the level of ZsGreen1 expression was tissue-specific. For example, ZsGreen1 was most abundantly produced in the salivary gland, moderately produced in the esophagus and lowly expressed in the stomach. Interestingly, expression of ZsGreen1 also differed within organ. For instance, levels were highest in the right ventricle compared with other chambers of the heart. The expression patterns of ZsGreen1 were similar between homozygous and hemizygous piglets. Ultimately, these results elucidate the tissuespecific activity of the CMV promoter in the neonatal pig.All rights reserved. No reuse allowed without permission.was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.

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Section: Identification Of Hemizygous and Homozygous Cmv-zsgreen1 Trasupporting

confidence: 70%

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confidence: 99%

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confidence: 99%

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A transgenic pig model expressing a ZsGreen1 reporter across an extensive array of tissues

Desaulniers

Cederberg

Carreiro

et al. 2018

Preprint

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“…Our results show near identical distribution profiles of developing thymocytes in the thymus and mature lymphocytes in the lymph node. Recent data shown preferential expression of the GFP in the exocrine cells of transgenic pigs, where the GFP transgene was placed under the strong cytomegalovirus (CMV) enhancer-promoter (Vasey et al 2009). …”

Section: Resultsmentioning

confidence: 99%

Characterisation of eGFP-transgenic BALB/c mouse strain established by lentiviral transgenesis

et al. 2009

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Lentiviral technology is a powerful tool for the creation of stable transgenic animals. However, uncertainties have remained whether constitutive promoters resist long-term silencing. We used concentrated HIV-1 based lentiviral vectors to create stable transgenic BALB/c mice by perivitelline injection. In our vectors eGFP expression was driven by the human EF1alpha promoter. The established transgenic animals were analyzed for eGFP expression by in vivo fluorescence imaging, PCR, histology and flow-cytometry. eGFP expression showed even distribution without mosaicism; however, tissue-dependent differences of eGFP expression were observed. Up to the sixth generation only one newborn showed eGFP inactivation. eGFP + transgenic bone marrow cells efficiently provided long-term haemopoietic repopulation in radiation chimeras, regenerating all bone marrow-derived lineages with eGFP + cells with distinct eGFP expression profiles. The established eGFP + BALB/c mouse strain is expected to be extremely useful in various immunological experiments.

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“…As such, and especially in non-rodent species where the standard methodology is either technically difficult or expensive, LV transgenesis is a very attractive addition to the tools available for transgene delivery. While the majority of publications to date have been with HIV-1-based vectors, alternative systems based on lentiviruses such as EIAV (Vasey et al 2009) or SIV (Hiripi et al 2010) are also used. However, both published concerns over transgene silencing (Hofmann et al 2006) and 'grapevine' comments on variable efficiency limit widespread uptake of the technology.…”

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confidence: 99%