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Clontech Laboratories, Inc. and RIKEN BioResource Research Center concluded license agreement on preservation and distribution of Fluorescent Proteins, DsRed2 and mCherry for academic use

BRC News

[RIKEN BRC]DNA Bank Mail News 2015.09.10.en


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RIKEN BioResource Center DNA Bank Mail News
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:: CONTENT :::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::
– The 74th Annual Meeting of the Japanese Cancer Association
– Pick Up Resource – Histac for visualization of acetylation of the Histon H4 –
– Focused Resource – Resources used in a recent paper –
– Genomic locus annotated BAC clones
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– The 74th Annual Meeting of the Japanese Cancer Association
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It is our pleasure to invite you to visit our booth in the event hall.

Date : October 8 (Thu) – October 10 (Sat), 2015 .
Venue : Nagoya Congress Center
The 74th Annual Meeting of the Japanese Cancer Association.

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– Pick Up Resource
– Histac for visualization of acetylation of the Histon H4 –
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Histac
– The fluorescent probe which detects acetylation of the histone H4 – Histone modification is thought to be one of the mechanisms of epigenetic gene regulation. However, probably because of lack of measures to observe living specimen, spatiotemporal dynamics analysis is in little progress.

Here, we offer fluorescent probe “Histac” deposited by Dr. Kazuki Sasaki (RIKEN Brain Science Institute) to detect global hyperacetylation of histone H4 in real-time living cells.

What is Histac?
Histac consists of an acetylation-binding domain, histone H4 as substrate, and the two fluorescent proteins.

Under the deacetylated conditions of the histon H4, CFP and Venus in the Histac molecule are closed and yield a yellow fluorescence (emission, 535 nm) by fluorescence resonance energy transfer (FRET). On the other hand, under the acetylated conditions, separation of the fluorescence proteins occurs by the structural change caused by the binding of the bromodomain to histone H4 in the Histac molecule, and yield a blue fluorescence (emission, 480 nm).

Observation of the acetylation of histone H4 by the Histaq requires no antibody reaction or fixation of cells. Histac allows successive observation of acetylation of the histone H4 in the introduced living animal cells.

In the two original papers, the authors observed fluorescence intensity ratio between CFP versus Venus (480 nm / 535 nm) progressively from 0 to 180 minutes after treatment of histone deacetylase inhibitor, trichostatin A (TSA).

The authors proved that Histaq as a powerful tool to show the dynamics of acetylation of histaq H4 in living cells.

References
1. Sasaki, K., et al. Real-time imaging of histone H4 hyperacetylation in living cells. PNAS, 106(38):16257-62, 2009.
2. Ito T, et al. Real-Time Imaging of Histone H4K12-Specific Acetylation Determines the Modes of Action of Histone Deacetylase and Bromodomain Inhibitors. Chemistry & Biology, 18(4):495-507, 2011.
3. RIKEN Research. Nov, 2009.

DNA Resource
Histac pcDNA3.1 (cat# RDB12840)
For the detection of acetylation at the 5th and 8th lysine residues of histone H4 by the BRDT bromodomain.

Histac-K12 pcDNA3.1 (cat# RDB12841)
For the detection of acetylation at the 12th lysine residues of histone H4 by the BRD2 bromodomain.

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– Focused Resource – Resources used in a recent paper –
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PRRX1 activates Notch signaling in the invasion of glioblastoma.
PRRX1 is involved in the invasion of glioblastoma. It is known that the activation of Notch, Wnt, and Hedgehog pathway also participate in the invasion of the glioblastoma. Sugiyama, M., et al performed a reporter assay using pGa981-6 (as for the Notch signal) [ cat. #RDB06776], 8×3’Gli-BS-delta51-LucII (as for the Hedgehog signal) [ cat. #RDB08061] to study activation of the pathways by PRRX1A expression.

Reference
Sugiyama, M., et al. Paired related homeobox 1 is associated with the invasive properties of glioblastoma cells. Oncol Rep.
33(3):1123-30, 2015.

DNA Resource
pGa981-6 (cat# RDB06776)
8×3’Gli-BS-delta51-LucII (cat# RDB08061)

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– Genomic locus annotated BAC clones
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RIKEN DNA Bank offers various BAC clones; Mouse (C57BL/6N, MSM/Ms strains), Rat (F344/Stm, LE/Stm, F344/Jcl, ACI/NJcl strains), Japanese macaque (M. fuscacta), and Drosophila flies (5 species, such as D. melanogaster).

BAC clones have been utilized to analyze the gene expression and to create transgenic (Tg) animals in the developmental biology and neuroscience fields. Mouse BAC clones have been also used to create Tg and knock-in animals for bioimaging technology using luciferases and fluorescent proteins.

You can search BAC clones through the BAC browser with gene symbols as a keyword, and confirm the physical location of BAC clones on the genome.

Please access the following address to search your BAC clones. We look forward to receiving your order.

2015.09.10