Bioluminescent system for dynamic imaging of cell and animal behavior

Hara-Miyauchi, Chikako; Tsuji, Osahiko; Hanyu, Aki; Okada, Seiji; Yasuda, Akimasa; Fukano, Takashi; Akazawa, Chihiro; Nakamura, Masaya; Imamura, Takeshi; Matsuzaki, Yuji; Okano, Hirotaka James; Miyawaki, Atsushi; Okano, Hideyuki

doi:10.1016/j.bbrc.2012.01.141

Cited by 66 publications

(74 citation statements)

References 21 publications

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“…The transplantation of neurospheres expressing ffLuc-cp156, a fusion protein of a fluorescent protein Venus and firefly luciferase, which had been introduced by lentiviral infection [23] was performed using a Hamilton Syringe with a stereotaxic injector, as described [24]. The needle of the Hamilton Syringe was inserted into the right striatum (2 mm lateral, 1 mm rostral to bregma; depth, 3 mm from dura) of 8-week-old female C57/b6j mice, and 3 ll of NSC suspension (2 Â 10 5 cells) was injected.…”

Section: Transplantationmentioning

confidence: 99%

Neural Stem Cells Directly Differentiated from Partially Reprogrammed Fibroblasts Rapidly Acquire Gliogenic Competency

et al. 2012

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Neural stem cells (NSCs) were directly induced from mouse fibroblasts using four reprogramming factors (Oct4, Sox2, Klf4, and cMyc) without the clonal isolation of induced pluripotent stem cells (iPSCs). These NSCs gave rise to both neurons and glial cells even at early passages, while early NSCs derived from clonal embryonic stem cells (ESCs)/ iPSCs differentiated mainly into neurons. Epidermal growth factor-dependent neurosphere cultivation efficiently propagated these gliogenic NSCs and eliminated residual pluripotent cells that could form teratomas in vivo. We concluded that these directly induced NSCs were derived from partially reprogrammed cells, because dissociated ESCs/ iPSCs did not form neurospheres in this culture condition. These NSCs differentiated into both neurons and glial cells in vivo after being transplanted intracranially into mouse striatum. NSCs could also be directly induced from adult human fibroblasts. The direct differentiation of partially reprogrammed cells may be useful for rapidly preparing NSCs with a strongly reduced propensity for tumorigenesis.

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Section: Transplantationmentioning

confidence: 99%

Neural Stem Cells Directly Differentiated from Partially Reprogrammed Fibroblasts Rapidly Acquire Gliogenic Competency

et al. 2012

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“…In ffLuc2-cp156, firefly luciferase (FLuc) was fused with a circularly permutated variant of Venus (cp156) [9], not for enhancement of light output by FRET, but rather for enhancement of protein stability and enzymatic activity. Indeed, transgenic ffLuc2-cp156 mice exhibited light output that was more than three orders of magnitude greater than that of transgenic FLuc mice.…”

Section: Luminescent Proteinsmentioning

confidence: 99%

Recent progress in luminescent proteins development

Saito

Nagai

2015

Current Opinion in Chemical Biology

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Bioimaging requires not only high sensitivity but also minimal invasiveness. Bioimaging using luminescent proteins is potentially free from problems such as photo-induced damage or an undesirable physical reaction to the sample, which are often caused by illumination with an external light required in fluorescence imaging. The recent development of several luminescent proteins and substrates have greatly improved the brightness of luminescence imaging, facilitating its application by many researchers. In this short review, we summarize recent advances in development of luminescent proteins, substrates, and indicators.

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“…1A). The ffLuc reporter gene is a fused gene of Venus and firefly luciferase sequences, and it allows both visualization of the expressing cells by fluorescence and quantification of the transcriptional activity by bioluminescence [19]. A 1-kb Msi1 promoter upstream of the Msi1 transcription start site was placed in the upstream portion of ffLuc.…”

Section: Msi1-6ie Reporter Gene Is Expressed In Msi1-positive Ns/pcs mentioning

confidence: 99%

Regulatory Factor X Transcription Factors Control Musashi1 Transcription in Mouse Neural Stem/Progenitor Cells

Kawase

Kuwako

Imai

et al. 2014

Stem Cells and Development

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The transcriptional regulation of neural stem/progenitor cells (NS/PCs) is of great interest in neural development and stem cell biology. The RNA-binding protein Musashi1 (Msi1), which is often employed as a marker for NS/ PCs, regulates Notch signaling to maintain NS/PCs in undifferentiated states by the translational repression of Numb expression. Considering these critical roles of Msi1 in the maintenance of NS/PCs, it is extremely important to elucidate the regulatory mechanisms by which Msi1 is selectively expressed in these cells. However, the mechanism regulating Msi1 transcription is unclear. We previously reported that the transcriptional regulatory region of Msi1 is located in the sixth intron of the Msi1 locus in NS/PCs, based on in vitro experiments. In the present study, we generated reporter transgenic mice for the sixth intronic Msi1 enhancer (Msi1-6IE), which show the reporter expression corresponding with endogenous Msi1-positive cells in developing and adult NS/PCs. We found that the core element responsible for this reporter gene activity includes palindromic Regulatory factor X (Rfx) binding sites and that Msi1-6IE was activated by Rfx. Rfx4, which was highly expressed in NS/PCs positive for the Msi1-6IE reporter, bound to this region, and both of the palindromic Rfx binding sites were required for the transactivation of Msi1-6IE. Furthermore, ectopic Rfx4 expression in the developing mouse cerebral cortex transactivates Msi1 expression in the intermediate zone. This study suggests that ciliogenic Rfx transcription factors regulate Msi1 expression through Msi1-6IE in NS/PCs.

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Bioluminescent system for dynamic imaging of cell and animal behavior

Cited by 66 publications

References 21 publications

Neural Stem Cells Directly Differentiated from Partially Reprogrammed Fibroblasts Rapidly Acquire Gliogenic Competency

Neural Stem Cells Directly Differentiated from Partially Reprogrammed Fibroblasts Rapidly Acquire Gliogenic Competency

Recent progress in luminescent proteins development

Regulatory Factor X Transcription Factors Control Musashi1 Transcription in Mouse Neural Stem/Progenitor Cells

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