Monomethylation of histone H4-lysine 20 is involved in chromosome structure and stability and is essential for mouse development.
AuthorsOda H, Okamoto I, Murphy N, Chu J, Price SM, Shen MM, Torres-Padilla ME, Heard E, Reinberg D,
JournalMol Cell Biol
PubMed ID19223465
'PR-Set7/Set8/KMT5A is the sole enzyme known to catalyze monomethylation of histone H4 lysine 20 (H4K20) and is present only in multicellular organisms that compact a large fraction of their DNA. We found that mouse embryos that are homozygous null mutants for the gene PR-Set7 display early embryonic lethality prior to ... More
Cell type specific applicability of 5-ethynyl-2'-deoxyuridine (EdU) for dynamic proliferation assessment in flow cytometry.
AuthorsDiermeier-Daucher S, Clarke ST, Hill D, Vollmann-Zwerenz A, Bradford JA, Brockhoff G,
JournalCytometry A
PubMed ID19235202
'Using the nucleoside analogue EdU (5-ethynyl-2''-deoxyuridine) for thymidine substitution instead of BrdU (5-bromo-2''-deoxyuridine) in cell proliferation assays has recently been proposed. However, the effect of EdU on cell viability, DNA synthesis, and cell cycle progression and consequently its usability for dynamic cell proliferation analysis in vitro has not been explored. ... More
Imaging and analysis of 3D tumor spheroids enriched for a cancer stem cell phenotype.
AuthorsRobertson FM, Ogasawara MA, Ye Z, Chu K, Pickei R, Debeb BG, Woodward WA, Hittelman WN, Cristofanilli M, Barsky SH,
JournalJ Biomol Screen
PubMed ID20639504
'Tumors that display a highly metastatic phenotype contain subpopulations of cells that display characteristics similar to embryonic stem cells. These cells exhibit the ability to undergo self-renewal; slowly replicate to retain a nucleoside analog label, leading to their definition as ' ... More
New strategy for synthesis of mercaptoundecahydrododecaborate derivatives via click chemistry: possible boron carriers and visualization in cells for neutron capture therapy.
AuthorsEl-Zaria ME, Nakamura H,
JournalInorg Chem
PubMed ID19928788
'A new method that utilizes the click cycloaddition reaction to functionalize B(12)H(11)SH(2-) (BSH) with organic molecules was investigated. S,S-Dipropargyl-SB(12)H(11)(-) (1) and S-propargyl-SB(12)H(11)(2-) (4) were prepared from [(CH(3))(4)N](2)B(12)H(11)SH and [(CH(3))(4)N](2)B(12)H(11)S(CH(2))(2)CN (2) with propargyl bromide, respectively. Compound 1 or 4 reacted with various azides with mediation by Cu(II) ascorbate to give the ... More
Cell cycle synchronization of Escherichia coli using the stringent response, with fluorescence labeling assays for DNA content and replication.
AuthorsFerullo DJ, Cooper DL, Moore HR, Lovett ST,
JournalMethods
PubMed ID19245839
'We describe a method for synchronization of the cell cycle in the bacterium Escherichia coli. Treatment of asynchronous cultures with the amino acid analog, dl-serine hydroxamate, induces the stringent response, with concomitant arrest of DNA replication at initiation. Following release of the stringent response, cells initiate DNA replication in synchrony, ... More
Click-iT assay with improved DNA distribution histograms.
AuthorsHamelik RM, Krishan A,
JournalCytometry A
PubMed ID19658154
'The Click-iT Assay developed and commercialized by Invitrogen is based on incorporation of a new 5-bromo-2''-deoxyuridine analog, 5-ethynyl-2''-deoxyuridine (EdU) into newly synthesized DNA and its recognition by azide dyes via a copper mediated ' ... More
EdU, a new thymidine analogue for labelling proliferating cells in the nervous system.
AuthorsChehrehasa F, Meedeniya AC, Dwyer P, Abrahamsen G, Mackay-Sim A,
JournalJ Neurosci Methods
PubMed ID18996411
Labelling and identifying proliferating cells is central to understanding neurogenesis and neural lineages in vivo and in vitro. We present here a novel thymidine analogue, ethynyl deoxyuridine (EdU) for labelling dividing cells, detected with a fluorescent azide which forms a covalent bond via the ... More
EdU incorporation is an alternative non-radioactive assay to [(3)H]thymidine uptake for in vitro measurement of mice T-cell proliferations.
AuthorsYu Y, Arora A, Min W, Roifman CM, Grunebaum E,
JournalJ Immunol Methods
PubMed ID19647746
RATIONALE: T lymphocyte proliferations can be measured by [(3)H]thymidine incorporation. However, many labs avoid this technique because of the need to use radioactive substrates. In addition, [(3)H]thymidine incorporation method does not permit simultaneous characterization of the proliferating cells. We developed the 5-ethynyl-2'-deoxyuridine (EdU) and Cu(I)-catalyzed cycloaddition ... More
AuthorsCronin SJ, Nehme NT, Limmer S, Liegeois S, Pospisilik JA, Schramek D, Leibbrandt A, Simoes Rde M, Gruber S, Puc U, Ebersberger I, Zoranovic T, Neely GG, von Haeseler A, Ferrandon D, Penninger JM,
JournalScience
PubMed ID19520911
Innate immunity represents the first line of defense in animals. We report a genome-wide in vivo Drosophila RNA interference screen to uncover genes involved in susceptibility or resistance to intestinal infection with the bacterium Serratia marcescens. We first employed whole-organism gene suppression, followed by tissue-specific silencing in gut epithelium or ... More
Detection of S-phase cell cycle progression using 5-ethynyl-2'-deoxyuridine incorporation with click chemistry, an alternative to using 5-bromo-2'-deoxyuridine antibodies.
The 5-bromo-2'-deoxyuridine (BrdU) labeling of cells followed by antibody staining has been the standard method for direct measurement of cells in the S-phase. Described is an improved method for the detection of S-phase cell cycle progression based upon the application of click chemistry, the copper(I)-catalyzed variant of the Huisgen [3+2] ... More
A rapid and robust assay for detection of S-phase cell cycle progression in plant cells and tissues by using ethynyl deoxyuridine.
AuthorsKotogány E, Dudits D, Horváth GV, Ayaydin F,
JournalPlant Methods
PubMed ID20181034
ABSTRACT: BACKGROUND: Progress in plant cell cycle research is highly dependent on reliable methods for detection of cells replicating DNA. Frequency of S-phase cells (cells in DNA synthesis phase) is a basic parameter in studies on the control of cell division cycle and the developmental events of plant cells. Here ... More
A rapid non-radioactive technique for measurement of repair synthesis in primary human fibroblasts by incorporation of ethynyl deoxyuridine (EdU).
AuthorsLimsirichaikul S, Niimi A, Fawcett H, Lehmann A, Yamashita S, Ogi T,
JournalNucleic Acids Res
PubMed ID19179371
Xeroderma pigmentosum (XP) is an autosomal recessive genetic disorder. Afflicted patients show extreme sun-sensitivity and skin cancer predisposition. XP is in most cases associated with deficient nucleotide excision repair (NER), which is the process responsible for removing photolesions from DNA. Measuring NER activity by nucleotide incorporation into repair patches, termed ... More
Chick embryo proliferation studies using EdU labeling.
AuthorsWarren M, Puskarczyk K, Chapman SC,
JournalDev Dyn
PubMed ID19253396
Cell proliferation studies are an important experimental tool. The most commonly used thymidine analogues, tritiated thymidine and bromodeoxyuridine (BrdU) label cells during S-phase. Both methods have significant drawbacks: low sensitivity in the case of tritiated thymidine and a denaturation step during BrdU detection that destroys most cellular epitopes, requiring careful ... More
METT-10, a putative methyltransferase, inhibits germ cell proliferative fate in Caenorhabditis elegans.
AuthorsDorsett M, Westlund B, Schedl T,
JournalGenetics
PubMed ID19596901
Germ-line stem cells are unique because they either self-renew through mitosis or, at a certain frequency, switch to meiosis and produce gametes. The switch from proliferation to meiosis is tightly regulated, and aberrations in switching result in either too little or too much proliferation. To understand the genetic basis of ... More
High-content, high-throughput analysis of cell cycle perturbations induced by the HSP90 inhibitor XL888.
AuthorsLyman SK, Crawley SC, Gong R, Adamkewicz JI, McGrath G, Chew JY, Choi J, Holst CR, Goon LH, Detmer SA, Vaclavikova J, Gerritsen ME, Blake RA,
JournalPLoS One
PubMed ID21408192
Many proteins that are dysregulated or mutated in cancer cells rely on the molecular chaperone HSP90 for their proper folding and activity, which has led to considerable interest in HSP90 as a cancer drug target. The diverse array of HSP90 client proteins encompasses oncogenic drivers, cell cycle components, and a ... More