Assays for DNA double-strand break repair by microhomology-based end-joining repair mechanisms.

DNA double stranded breaks (DSBs) are one of the most deleterious types of DNA lesions. The main pathways responsible for repairing these breaks in eukaryotic cells are homologous recombination (HR) and non-homologous end-joining (NHEJ). However, a third group of still poorly characterized DSB repair pathways, collectively termed microhomology-mediated end-joining (MMEJ), relies on short homologies for the end-joining process. Here, we constructed GFP reporter assays to characterize and distinguish MMEJ variant pathways, namely the simple MMEJ and the DNA synthesis-dependent (SD)-MMEJ mechanisms. Transfection of these assay vectors in Chinese hamster ovary (CHO) cells and characterization of the repaired DNA sequences indicated that while simple MMEJ is able to mediate relatively efficient DSB repair if longer microhomologies are present, the majority of DSBs were repaired using the highly error-prone SD-MMEJ pathway. To validate the involvement of DNA synthesis in the repair process, siRNA knock-down of different genes proposed to play a role in MMEJ were performed, revealing that the knock-down of DNA polymerase θ inhibited DNA end resection and repair through simple MMEJ, thus favoring the other repair pathway. Overall, we conclude that this approach provides a convenient assay to study MMEJ-related DNA repair pathways

Application of large area SiPMs for the readout of a plastic scintillator based timing detector

In this study an array of eight 6 mm x 6 mm area SiPMs was coupled to the end of a long plastic scintillator counter which was exposed to a 2.5 GeV/c muon beam at the CERN PS. Timing characteristics of bars with dimensions 150 cm x 6 cm x 1 cm and 120 cm x 11 cm x 2.5 cm have been studied. An 8-channel SiPM anode readout ASIC (MUSIC R1) based on a novel low input impedance current conveyor has been used to read out and amplify SiPMs independently and sum the signals at the end. Prospects for applications in large-scale particle physics detectors with timing resolution below 100 ps are provided in light of the results

Application of large area SiPMs for the readout of a plastic scintillator based timing detector

In this study an array of eight 6 mm x 6 mm area SiPMs was coupled to the end of a long plastic scintillator counter which was exposed to a 2.5 GeV/c muon beam at the CERN PS. Timing characteristics of bars with dimensions 150 cm x 6 cm x 1 cm and 120 cm x 11 cm x 2.5 cm have been studied. An 8-channel SiPM anode readout ASIC (MUSIC R1) based on a novel low input impedance current conveyor has been used to read out and amplify SiPMs independently and sum the signals at the end. Prospects for applications in large-scale particle physics detectors with timing resolution below 100 ps are provided in light of the results

MAR-Mediated Dystrophin Expression in Mesoangioblasts for Duchenne Muscular Dystrophy Cell Therapy

A cornerstone of autologous cell therapy for Duchenne muscular dystrophy is the engineering of suitable cells to express dystrophin in a stable fashion upon differentiation to muscle fibers. Most viral transduction methods are typically restricted to the expression of truncated recombinant dystrophin derivatives and by the risk of insertional mutagenesis, while non-viral vectors often suffer from inefficient transfer, expression and/or silencing

Light-ion Production And Fission Studies Using The Medley Facility At Tsl

oS(FNDA2006)001 © Copyright owned by the author(s) under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike Licence

Steady-state expression of self-regulated genes

Motivation: Regulatory gene networks contain generic modules such as feedback loops that are essential for the regulation of many biological functions. The study of the stochastic mechanisms of gene regulation is instrumental for the understanding of how cells maintain their expression at levels commensurate with their biological role, as well as to engineer gene expression switches of appropriate behavior. The lack of precise knowledge on the steady-state distribution of gene expression requires the use of Gillespie algorithms and Monte-Carlo approximations. Methodology: In this study, we provide new exact formulas and efficient numerical algorithms for computing/modeling the steady-state of a class of self-regulated genes, and we use it to model/compute the stochastic expression of a gene of interest in an engineered network introduced in mammalian cells. The behavior of the genetic network is then analyzed experimentally in living cells. Results: Stochastic models often reveal counter-intuitive experimental behaviors, and we find that this genetic architecture displays a unimodal behavior in mammalian cells, which was unexpected given its known bimodal response in unicellular organisms. We provide a molecular rationale for this behavior, and we implement it in the mathematical picture to explain the experimental results obtained from this network. Contact: [email protected], [email protected] Supplementary information: Supplementary data are available at Bioinformatics onlin

Light-ion production in the interaction of 96 MeV neutrons with oxygen

Double-differential cross sections for light-ion (p, d, t, He-3 and alpha) production in oxygen, induced by 96 MeV neutrons are reported. Energy spectra are measured at eight laboratory angles from 20 degrees to 160 degrees in steps of 20 degrees. Procedures for data taking and data reduction are presented. Deduced energy-differential and production cross sections are reported. Experimental cross sections are compared to theoretical reaction model calculations and experimental data at lower neutron energies in the literature. The measured proton data agree reasonably well with the results of the model calculations, whereas the agreement for the other particles is less convincing. The measured production cross sections for protons, deuterons, tritons and alpha particles support the trends suggested by data at lower energies.Comment: 21 pages, 13 figures, submitted to Phys. Rev.

Light-ion production in the interaction of 96 MeV neutrons with silicon

Double-differential cross sections for light-ion (p, d, t, He-3 and alpha) production in silicon, induced by 96 MeV neutrons are reported. Energy spectra are measured at eight laboratory angles, ranging from 20 degrees to 160 degrees in steps of 20 degrees. Procedures for data taking and data reduction are presented. Deduced energy-differential, angle-differential and production cross sections are reported. Experimental cross sections are compared to theoretical reaction model calculations and experimental data in the literature.Comment: 23 pages, 10 figures, added wrap-around correction (see section 4.3) leading to changed cross-sections and figures, accepted Phys. Rev.

Extracellular deposition of matrilin-2 controls the timing of the myogenic program during muscle regeneration.

Here, we identify a role for the matrilin-2 (Matn2) extracellular matrix protein in controlling the early stages of myogenic differentiation. We observed Matn2 deposition around proliferating, differentiating and fusing myoblasts in culture and during muscle regeneration in vivo. Silencing of Matn2 delayed the expression of the Cdk inhibitor p21 and of the myogenic genes Nfix, MyoD and Myog, explaining the retarded cell cycle exit and myoblast differentiation. Rescue of Matn2 expression restored differentiation and the expression of p21 and of the myogenic genes. TGF-β1 inhibited myogenic differentiation at least in part by repressing Matn2 expression, which inhibited the onset of a positive-feedback loop whereby Matn2 and Nfix activate the expression of one another and activate myoblast differentiation. In vivo, myoblast cell cycle arrest and muscle regeneration was delayed in Matn2(-/-) relative to wild-type mice. The expression levels of Trf3 and myogenic genes were robustly reduced in Matn2(-/-) fetal limbs and in differentiating primary myoblast cultures, establishing Matn2 as a key modulator of the regulatory cascade that initiates terminal myogenic differentiation. Our data thus identify Matn2 as a crucial component of a genetic switch that modulates the onset of tissue repair