センサー&インジケーター

Sensor & Visualization

ATP measurement

  • QUEEN
    Diversity in ATP concentrations in a single bacterial cell population revealed by quantitative single-cell imaging.
    Yaginuma, H., Kawai, S., Tabata, K.V., Tomiyama, K., Kakizuka, A., Komatsuzaki, T., Noji, H., Imamura, H.
    Sci. Rep. 4: 6522 (2014). PMID 25283467.

Autophagy indicator

  • Chaperone mediated autophagy activity by the GAPDH-HT indicator by Dr. Takahiro Seki’s lab as well as fluorescent protein probes of LC3B accumulation developed by Dr. Noboru Mizushima’s lab, Dr. Itaru Hamachi’s lab and Dr. Keiji Kimura’s lab are available.

Bilirubin インジケーター UnaG, BReleaCa

  • A dual-ligand-modulable fluorescent protein based on UnaG and calmodulin.
    Shitashima, Y., Shimozawa, T., Asahi, T., Miyawaki, A.
    Biochem. Biophys. Res. Commun. 496 (3): 872-879 (2018). PubMed PMID 29395087.

    Catalog no. Name of resource Descriptiion
    RDB15908 pRSETB-BReleaCa Bacteria expression vector of UnaG/CaM hybrid protein, BReleaCa, capable of binding to bilirubin and calcium.
    Catalog no. Name of resource Descriptiion
    RDB15909 pcDNA3-BReleaCa Expression vector of UnaG/CaM hybrid protein, BReleaCa, capable of binding to bilirubin and calcium.
  • Continuous de novo biosynthesis of haem and its rapid turnover to bilirubin are necessary for cytoprotection against cell damage.
    Takeda, T.A., Mu, A., Tai, T.T., Kitajima, S., Taketani, S.
    Sci. Rep. 5: 10488 (2015). PubMed PMID 25990790.

    Catalog no. Name of resource Descriptiion
    RDB13504 pET-His UNAG Expression clone of eel UnaG in E.coli.
  • A bilirubin-inducible fluorescent protein from eel muscle.
    Kumagai, A., Ando, R., Miyatake, H., Greimel, P., Kobayashi, T., Hirabayashi, Y., Shimogori, T., Miyawaki, A.
    Cell 153 (7): 1602-1611(2013). PubMed PMID 23768684.

    Catalog no. Name of resource Descriptiion
    RDB15703 UnaG/pcDNA3-FLAG Expression vector of eel UnaG, bilirubin-inducible fluorescent protein.
    Catalog no. Name of resource Descriptiion
    RDB15704 UnaG/pGEX-2T xpression vector of eel UnaG, bilirubin-inducible fluorescent protein.

cAMPインジケーター

  • Extracellular calcium influx activates adenylate cyclase 1 and potentiates insulin secretion in MIN6 cells.
    Kitaguchi, T., Oya, M., Wada, Y., Tsuboi, T., Miyawaki, A.
    Biochem. J. 450 (2): 365-373 (2013). PubMed PMID 23282092.

    Catalog no. Name of resource Descriptiion
    RDB15247 Flamindo/pcDNA3 Expression vector of yellow fluorescent biosensor for cAMP, Flamindo.

Cell cycle indicator Fucci

  • To monitor cell cycle progression in living cells, cell cycle indicator Fucci probes deveoped by Dr. Atsushi Miyawaki’s lab are available.

Caspase activity indicator

  • A high-throughput method for development of FRET-based indicators for proteolysis.
    Nagai, T., Miyawaki, A.
    Biochem. Biophys. Res. Commun. 319 (1): 72-77 (2004). PubMed PMID 15158444.

    Catalog no. Name of resource Descriptiion
    RDB15705 SCAT3.1/pcDNA4 HMB Expression vector of FRET based apoptosis probe.
  • Cyan-emitting and orange-emitting fluorescent proteins as a donor/acceptor pair for fluorescence resonance energy transfer.
    Karasawa, S., Araki, T., Nagai, T., Mizuno, H., Miyawaki, A.
    Biochem. J. 381 (Pt 1): 307-312 (2004). PubMed PMID 15065984.

    Catalog no. Name of resource Descriptiion
    RDB15240 MiCy-DEVD-mKO1/pCS2 Expression vector of FRET based caspase3 activity monitoring probe.

Calcium-ion sensor

  • Fluorescent protein-based Ca2+ sensors, G-CaMPs, Yellow Cameleons and Pericams developed by Dr. Jin-ichi Nakai’s lab and Dr. Atsushi Miyawaki’s lab, which are composed with calmodulin, fluorescent protein and M13 peptide (CaM binding domain of myosin light chain kinase), are designated to visualize intracellular [Ca2+] dynamics.
  • G-CaMP
  • Pericam
  • Yellow Cameleon

細胞内分子混雑インジケーター GimRET

  • Dependence of fluorescent protein brightness on protein concentration in solution and enhancement of it.
    Morikawa, J.T., Fujita, H., Kitamura, A., Horio, T., Yamamoto, J., Kinjo, M., Sasaki, A., Machiyama, H., Yoshizawa, K., Ichimura, T., Imada, K., Nagai, T., Watanabe, M.T.
    Sci. Rep. 6: 22342, 2016. PubMed PMID: 26956628

    Catalog no. Name of resource Descriptiion
    RDB14203 pPAL7_GimRET (CFP-YFP1G/pPAL7) Expression vector of GimRET (Glycine inserted mutant FRET probe) for E. coli.
    RDB14204 pGimRET (CFP-YFP1G/pECFP) Expression vector of GimRET (Glycine inserted mutant FRET probe) for mammalian cells.

pH センサー

  • pH に応じて青色 (456nm) から緑色 (507nm) に変化する蛍光を有する ウミサボテン Cavernularia obesa 由来のGFP です。哺乳動物細胞内における pH センサーとして活用できます。
    Ogoh, K. et al. Dual-color-emitting green fluorescent protein from the sea cactus Cavernularia obesa and its use as a pH indicator for fluorescence microscopy. Luminescence 28 (4): 582-591, 2013.

    Catalog no. Name of clone Characteristic misc.
    RDB14368 pCoGFP-wt wild type
    362nm で励起した各種 pH 感受性 GFP の pH4 – 11 における青色から緑色への蛍光推移をイメージしたイラスト。詳しくは、Ogoh, K. et al. Luminescence 28 (4): 582-591, 2013 の Figure 6 をご参考ください。
    RDB14369 pCoGFP-mam From blue to green at pH 5 – 6
    RDB14370 pCoGFP-V0 From blue to green at pH 5 – 6
    RDB14371 pCoGFP-V1 From blue to green at pH 6 – 7
    RDB14372 pCoGFP-V2 From blue to green at pH 7 – 8
    RDB14373 pCoGFP-V3 From blue to green at pH 9 – 10
    RDB14374 pCoGFP-V4 From blue to green at pH 9 – 10

RAインジケーター

  • Visualization of an endogenous retinoic acid gradient across embryonic development.
    Shimozono, S., Iimura, T., Kitaguchi, T., Higashijima, S., Miyawaki, A.
    Nature 496 (7445): 363-366 (2013). PubMed PMID 23563268.

    Catalog no. Name of resource Descriptiion
    RDB15249 GEPRA-B Genetically encoded indicator for RA, GEPRA, having ligand binding domain from the RAR-beta.
    RDB15250 GEPRA-G Genetically encoded indicator for RA, GEPRA, having ligand binding domain from the RAR-gamma.
    RDB15251 GEPRA-AA Genetically encoded indicator for RA, GEPRA. Low-affinity verson of the GEPRA-B.
    RDB15252 GEPRA-B/ pT2KXIGdeltain Expression vector of genetically encoded indicator for RA, GEPRA. For generation of transgenic zebrafish.
    RDB15253 GEPRA-G/ pT2KXIGdeltain Expression vector of genetically encoded indicator for RA, GEPRA. For generation of transgenic zebrafish.
    RDB15254 GEPRA-AA/ pT2KXIGdeltain Expression vector of genetically encoded indicator for RA, GEPRA. For generation of transgenic zebrafish. Low-affinity verson of the GEPRA-B.

Rab11: active Rab11 visualization

  • RBD11, a bioengineered Rab11-binding module for visualizing and analyzing endogenous Rab11.
    Osaki F, Matsui T, Hiragi S, Homma Y, Fukuda M.
    J. Cell Sci. 134 (7): jcs257311 (2021). PubMed PMID 33712449.

    Catalog no. Name of resource Descriptiion
    RDB18788 pMRX-puro-EGFP-RBD11 Retroviral expression vector for visualization of active Rab11: mouse Rab11-specific binding domain (RBD11) tagged with EGFP.
    RDB18789 pMRX-puro-EGFP-RBD11-mut Retroviral expression vector of negative control for visualization of active Rab11: mouse Rab11-specific binding domain (RBD11) mutant tagged with EGFP.
    RDB18790 pMRX-puro-EGFP-2xRBD11 Retroviral expression vector of negative control for visualization of active Rab11: mouse Rab11-specific binding domain (RBD11) mutant tagged with EGFP.
    RDB18791 pMRX-puro-EGFP-2xRBD11-mut Retroviral expression vector of negative control for visualization of active Rab11: mouse Rab11-specific binding domain (RBD11) mutant tagged with EGFP.
    RDB18792 pMRX-puro-EGFP-FM-RBD11 Retroviral expression vector of active Rab11 trapper: FM-tagged mouse Rab11-specific binding domain (RBD 11) tagged with EGFP.
    RDB18793 pMRX-puro-EGFP-FM-RBD11-mut Retroviral expression vector of negative control of active Rab11 trapper: FM-tagged mouse Rab11-specific binding domain (RBD 11) mutant tagged with EGFP.

Sphingolipid marker

  • Lipid rafts are small lipid domains on the cell membrane and are thought to play an important role in signal transduction, endocytosis and more. We provide fluorescent probes for sphingomyelin and cholesterol lipid domains.
  • Nakanori: sphingomyelin and cholesterol lipid domain (lipid raft)
  • D4 toxin: cholesterol rich domain
  • lysenin: sphingomyelin

Voltageインジケーター

  • Improving membrane voltage measurements using FRET with new fluorescent proteins.
    Tsutsui, H., Karasawa, S., Okamura, Y., Miyawaki, A.
    Nat. Methods 5 (8): 683-685 (2008). PubMed PMID 18622396.

    Catalog no. Name of resource Descriptiion
    RDB15262 Mermaid Voltage indicator Mermaid.
  • Improved detection of electrical activity with a voltage probe based on a voltage-sensing phosphatase.
    Tsutsui, H., Jinno, Y., Tomita, A., Niino, Y., Yamada, Y., Mikoshiba, K., Miyawaki, A., Okamura, Y.
    J. Physiol. 591 (18): 4427-4437 (2013). PubMed PMID 23836686.

    Catalog no. Name of resource Descriptiion
    RDB15263 Mermaid2 Voltage probe Mermaid2.

Notch signaling reporter

  • The pRBS-EGFP and RBP-J-Venus expression clones deposited by Dr. Makoto Mark Taketo and Dr. Kenji Tanigaki, respectively, allow you monitoring the state of activation of the Notch signaling by fluorescence in living cells.

Epigenetics reporter

  • Visualization of histone acetylation: The Histac fluorescent probes deposited by Dr. Kazuki Sasaki allow you monitoring the state of activity of acetylation of histone H4 by fluorescence in living cells.
  • Visualization of methylated DNA: The EGFP-MBD-nls protein recognizes the methylated DNA and you can follow status of the DNA methylation in situ under physiological conditions using the pEGFP-MBD-nls expression clone.

細胞内小器官/局在マーカー

References


蛍光タンパク質リソース

  • ウミサボテン由来耐熱性GFP
  • Azami Green (AG, hmAG1, hmAG407)
  • cjBlue (cjBlue, cjBlue Y64L)
  • Cy11.5
  • Dronpa-Green (Dronpa, Dronpa2, Dronpa3, 22G)
  • Kaede
  • Keima-red (dKeima, dKeima570, mKeima, tdKeima)
  • Kikume Green-Red (KikGR, mKikGR, mKikGR13.2, Xpa)
  • Kusabira Green Orange (mK-GO)
  • Kusabira Orange (KO1, hmKO1, hmKO2, hmKO-K)
  • Midoriishi-Cyan (MiCy, mMiCy1)
  • Venus (Venus, mVenus, cp49Venus, cp145Venus, cp157Venus, cp173Venus, cp195Venus, cp229Venus)
  • Destabilized FPs
  • Knock in markers with CRISPR/Cas9 genome editing

Luminescent protein

  • 高発光、緑色黄色赤色のルシフェラーゼ
    • 一般的な北米産ホタル由来のルシフェラーゼと比較して、2 倍以上、最大で 4倍程の明るい発光強度が得られる日本産、ならびにマレーシア産ホタル由来のルシフェラーゼ遺伝子です。オリンパス株式会社 小江克典先生らによって開発されました。基質である D- ルシフェリンとの反応によって高い発光強度を得られることが確認されています。
  • Akaluc -新規開発の赤色発光のルシフェラーゼ
    • AkaBLIシステムは生きた動物個体深部を非侵襲的に観察できる人工生物発生システムです。理研脳神経科学研究センターの宮脇 敦史先生、岩野 智先生、電気通信大学大学の牧 昌次郎先生らのグループにより開発されました。組織透過性の向上した人工基質AkaLumineとAkaLumineに合わせて開発した人工酵素Akalucで構成されています。AkaBLIの発光シグナルの強度は、従来と比べ100~1000倍です。
  • Nano lantern: ナノ ランタン
    • ウミシイタケルシフェラーゼと蛍光タンパク質を細胞内や核内等で物理的に隣接させ、ルシフェラーゼによる化学発光を光源とすることで、蛍光タンパク質の観察を可能にするリサーチツールです。蛍光タンパク質には、青緑色黄緑色橙色の3色があり、理化学研究所生命機能科学研究センターの岡田康志先生、高井啓先生、大阪大学産業科学研究所の永井健治先生等によって開発されました。

 

(GRP0058j 2019.01.24 T.M.)

2021.09.09



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