Cytoplasmic β‐actin promoter produces germ cell and preimplantation embryonic transgene expression

Sands, Arthur; Hansen, Thomas N.; DeMayo, Francesco J.; Stanley, L. A.; Xin, Lei; Schwartz, Robert J.

doi:10.1002/mrd.1080340202

Cited by 20 publications

(7 citation statements)

References 46 publications

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“…To test this hypothesis, we linked the VP16 gene to a human ␤-actin gene promoter to generate transgenic mice. As the human ␤-actin promoter has been shown to be active in embryos as early as the 2-cell stage (Nilsson and Lendahl, 1993), it was expected to drive VP16 expression ubiquitously throughout embryogenesis (Sands et al, 1993). The structure of the ␤-actin-VP16 transgene (construct 1) is depicted in Figure 1.…”

Section: Resultsmentioning

confidence: 99%

Herpes simplex virus transcriptional activator VP16 is detrimental to preimplantation development in mice

2000

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The herpes simplex virus transactivator protein VP16 is frequently used to regulate gene expression in several experimental systems, including transgenic mice. It has been suggested that high levels of VP16 expression in mice may be lethal. In order to systematically address this issue, we linked the VP16 gene to promoters that are active early and in a variety of tissues throughout development, such as the human β‐actin promoter or the rat nestin gene enhancer. VP16 expression was assayed using a LacZ reporter gene linked to a VP16‐responsive immediate early gene promoter. We show here that expression of VP16 at high levels is detrimental to pre‐implantation development. By culturing embryos in vitro, we demonstrate that this effect is exerted at the transition from the 2‐cell to the 4‐cell stage, reducing survival to the blastocyst stage dramatically. On the other hand, transgenic mice expressing VP16 transgenes at postimplantation stages are viable. These results suggest a differential sensitivity to VP16 expression in different cell types and stages of development. The reduction of embryo survival by VP16 implicates herpes virus infection as a potential cause of infertility. Mol. Reprod. Dev. 55:37–46, 2000. © 2000 Wiley‐Liss, Inc.

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Section: Resultsmentioning

confidence: 99%

Herpes simplex virus transcriptional activator VP16 is detrimental to preimplantation development in mice

2000

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“…Hence, mouse strains that express a marker gene in all tissues and cells throughout development are highly desirable. Several short promoters potentially capable of driving widespread tissue expression of marker genes include SV40 (Furth et al, 1991;Takeda and Toyoda, 1991), CMV (Furth et al, 1991), chicken b-actin (Sands et al, 1993), as well as a combination of CMV and b-actin (Okabe et al, 1997). Although most of these reporter lines show transgene expression in many tissues, the transgene is often not universally and reproducibly expressed within tissues and between animals.…”

Section: Introductionmentioning

confidence: 99%

Ubiquitous and uniform in vivo fluorescence in ROSA26‐EGFP BAC transgenic mice

Giel–Moloney¹,

Krause²,

Chen³

et al. 2007

Genesis

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Transplantation studies and cell lineage analyses require the ability to explicitly distinguish morphologically identical cells that have an identifiable marker indicating their origin in vivo. Several reporter mouse strains have been generated for such studies, but pancellular detection of the marker in all tissues has not been achieved. In this report, we describe the generation of transgenic mice that express enhanced green fluorescent protein (EGFP) under control of a 187 kb bacterial artificial chromosome (BAC) containing the murine ROSA26 locus, and show several advantages over existing EGFP reporter lines. It is demonstrated that EGFP is ubiquitously and reproducibly expressed from the murine BAC transgene in all organs and tissues analyzed, including the hematolymphoid compartment. Using this new reporter strain in hematopoietic cell transplantation studies, it is demonstrated that leukocytes in recipients maintain uniform transgene expression and are easily distinguished by flow cytometric analysis of live cells. The results suggest that the ROSA26 BAC is an efficient strategy for expressing complex transgene cassettes in vivo.

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“…Panjang sekuens hasil amplifikasi PCR mirip dengan panjang sekuens hasil sekuensing, yaitu 2,2 kb. Panjang sekuens arah forward 1.262 kb, arah reverse 1.202 kb, dan bagian sekuens yang sama sepanjang 248 bp M24113, medaka S74868 and nile tilipia AY116536 al., 1989;Sands et al, 1993). Dengan demikian dapat disimpulkan bahwa sekuens promoter β-aktin ikan gurami adalah konserf secara evolusi (evolutionary conserved).…”

Section: Figure 2 Genomic Sequence Of Gouramy β-Actin From Forward Dunclassified

KLONING PROMOTER b-AKTIN DARI IKAN GURAMI (Osphronemus gouramy)

Nugroho

Alimuddin

Kristanto

et al. 2016

Jurnal Riset Akuakultur

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Penelitian ini dilakukan untuk mengisolasi promoter b-aktin (ggBA) dari ikan gurami (Osphronemus gouramy). Sekuens promoter ggBA diisolasi menggunakan metode “degenerate” PCR. Sekuensing dilakukan menggunakan mesin ABI PRISM 3100-Avant. Analisis sekuens menggunakan program BLAST, GENETYX versi 7 dan TFBind. Panjang sekuens DNA hasil kloning adalah sekitar 2,2 kb. Analisis BLAST menunjukkan bahwa sekuens DNA hasil kloning memiliki kemiripan dengan sekuens gen b-aktin ikan yang ada di Bank Gen. Sekuens hasil kloning memiliki faktor transkripsi yang konserf untuk promoter b-aktin, yaitu: CCAAT, CC(A/T)6GG, dan boks TATA. Posisi faktor transkripsi tersebut juga mirip dengan yang dimiliki oleh promoter b-aktin dari ikan yang ada di Bank Gen. Dengan demikian dapat disimpulkan bahwa fragmen DNA hasil amplifikasi PCR tersebut merupakan sekuens promoter b-aktin ikan gurami.The research was aimed to isolate b-actin promoter of gouramy (ggBA). Sequence of ggBA promoter was isolated using “degenerate” PCR. Sequencing was conducted using ABI PRISM 3100-Avant machine. The sequencing analysis was done using BLAST, GENETYX ver 7 and TFBind programs. The sequencing length of DNA clone result was around 2.2 kb. BLAST analysis showed that sequences from cloned DNA had the resemblence to those of b-actin sequences in GenBank database. The cloned sequences had transcript factors which followed b-actin promoter namely CCAAT, CC(A/T)6GG and TATA box.

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Cytoplasmic β‐actin promoter produces germ cell and preimplantation embryonic transgene expression

Cited by 20 publications

References 46 publications

Herpes simplex virus transcriptional activator VP16 is detrimental to preimplantation development in mice

Herpes simplex virus transcriptional activator VP16 is detrimental to preimplantation development in mice

Ubiquitous and uniform in vivo fluorescence in ROSA26‐EGFP BAC transgenic mice

KLONING PROMOTER b-AKTIN DARI IKAN GURAMI (Osphronemus gouramy)

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