Improved genome editing in human cell lines using the CRISPR method

The Cas9/CRISPR system has become a popular choice for genome editing. In this system, binding of a single guide (sg) RNA to a cognate genomic sequence enables the Cas9 nuclease to induce a double-strand break at that locus. This break is next repaired by an error-prone mechanism, leading to mutation and gene disruption. In this study we describe a range of refinements of the method, including stable cell lines expressing Cas9, and a PCR based protocol for the generation of the sgRNA. We also describe a simple methodology that allows both elimination of Cas9 from cells after gene disruption and re-introduction of the disrupted gene. This advance enables easy assessment of the off target effects associated with gene disruption, as well as phenotype-based structure-function analysis. In our study, we used the Fan1 DNA repair gene as control in these experiments. Cas9/CRISPR-mediated Fan1 disruption occurred at frequencies of around 29%, and resulted in the anticipated spectrum of genotoxin hypersensitivity, which was rescued by re-introduction of Fan1

Proton-induced magnetic order in carbon: SQUID measurements

In this work we have studied systematically the changes in the magnetic behavior of highly oriented pyrolytic graphite (HOPG) samples after proton irradiation in the MeV energy range. Superconducting quantum interferometer device (SQUID) results obtained from samples with thousands of localized spots of micrometer size as well on samples irradiated with a broad beam confirm previously reported results. Both, the para- and ferromagnetic contributions depend strongly on the irradiation details. The results indicate that the magnetic moment at saturation of spots of micrometer size is of the order of $10^{-10}$ emu.Comment: Invited contribution at ICACS2006 to be published in Nucl. Instr. and Meth. B. 8 pages and 6 figure

Fingerprints of Spin-Orbital Physics in Crystalline O2_2

The alkali hyperoxide KO$_2$ is a molecular analog of strongly-correlated systems, comprising of orbitally degenerate magnetic O$_2^-$ ions. Using first-principles electronic structure calculations, we set up an effective spin-orbital model for the low-energy \textit{molecular} orbitals and argue that many anomalous properties of KO$_2$ replicate the status of its orbital system in various temperature regimes.Comment: 4 pages, 2 figures, 1 tabl

Induced Magnetic Ordering by Proton Irradiation in Graphite

We provide evidence that proton irradiation of energy 2.25 MeV on highly-oriented pyrolytic graphite samples triggers ferro- or ferrimagnetism. Measurements performed with a superconducting quantum interferometer device (SQUID) and magnetic force microscopy (MFM) reveal that the magnetic ordering is stable at room temperature.Comment: 3 Figure

First-Principles Study of Substitutional Metal Impurities in Graphene: Structural, Electronic and Magnetic Properties

We present a theoretical study using density functional calculations of the structural, electronic and magnetic properties of 3d transition metal, noble metal and Zn atoms interacting with carbon monovacancies in graphene. We pay special attention to the electronic and magnetic properties of these substitutional impurities and found that they can be fully understood using a simple model based on the hybridization between the states of the metal atom, particularly the d shell, and the defect levels associated with an unreconstructed D3h carbon vacancy. We identify three different regimes associated with the occupation of different carbon-metal hybridized electronic levels: (i) bonding states are completely filled for Sc and Ti, and these impurities are non-magnetic; (ii) the non-bonding d shell is partially occupied for V, Cr and Mn and, correspondingly, these impurties present large and localized spin moments; (iii) antibonding states with increasing carbon character are progressively filled for Co, Ni, the noble metals and Zn. The spin moments of these impurities oscillate between 0 and 1 Bohr magnetons and are increasingly delocalized. The substitutional Zn suffers a Jahn-Teller-like distortion from the C3v symmetry and, as a consequence, has a zero spin moment. Fe occupies a distinct position at the border between regimes (ii) and (iii) and shows a more complex behavior: while is non-magnetic at the level of GGA calculations, its spin moment can be switched on using GGA+U calculations with moderate values of the U parameter.Comment: 13 figures, 4 tables. Submitted to Phys. Rev. B on September 26th, 200

Interaction Between Superconducting and Ferromagnetic Order Parameters in Graphite-Sulfur Composites

The superconductivity of graphite-sulfur composites is highly anisotropic and associated with the graphite planes. The superconducting state coexists with the ferromagnetism of pure graphite, and a continuous crossover from superconducting to ferromagnetic-like behavior could be achieved by increasing the magnetic field or the temperature. The angular dependence of the magnetic moment m(alpha) provides evidence for an interaction between the ferromagnetic and the superconducting order parameters.Comment: 11 pages, 4 figures, to be published in Phys. Rev.

КЛИНИКО-ПАТОГЕНЕТИЧЕСКОЕ ОБОСНОВАНИЕ И ЭФФЕКТИВНОСТЬ ПРИМЕНЕНИЯ ВНУТРИВЕННОГО ИММУНОГЛОБУЛИНА У ДЕТЕЙ С БАКТЕРИАЛЬНЫМИ ГНОЙНЫМИ МЕНИНГИТАМИ

The article presents data on a study of the effectiveness of intravenous application of immunoglobulin in children with purulent bacterial meningitis of different etiology progressing with pronounced cephaledema. According to our data, the reason for substitution of the immunoglobulin deficiency in patients with purulent bacterial meningitis is clinical syndromes, expressed in points and laboratory indicators of the level of IgG in blood and cerebrospinal fluid of children. The findings give evidence of the efficacy of Intravenous Immunoglobulin application at severe purulent bacterial meningitis accompanied by pronounced immune deficiency, manifested in insufficient synthesis of IgG.В статье представлены данные об изучении эффективности использовании внутривенного иммуноглобулина (ВИГ) у детей, больных бактериальными гнойными менингитами (БГМ) различной этиологии, протекающими с выраженным отеком головного мозга. Основанием для замещения иммунодефицита иммуноглобулинов у больных БГМ, по нашим данным, являлись клинические синдромы, выраженные в баллах и лабораторные показатели уровня IgG в крови, ликворе детей. Полученные данные свидетельствуют об эффективности применения ВИГа при БГМ, протекающих в тяжелой степени тяжести, которые сопровождаются выраженной иммунологической недостаточностью, проявляющейся недостаточным синтезом IgG

Magnetic properties of carbon phases synthesized using high pressure-high temperature treatment

Two sets of samples were synthesized at 3.5 GPa near the point of C60 cage collapse at different annealing times. A clear structural transformation from mixture of C60 polymeric phases to graphite-like hard carbon phase was confirmed by X-ray diffraction and Raman spectroscopy. Magnetic force microscopy and superconducting quantum interference device were used to characterize the magnetic properties of the synthesized samples. We found that the sample preparation conditions used in this study are not suitable to produce bulk magnetic carbon.Comment: 26 pages, 7 figure

High-temperature ferromagnetism of spsp electrons in narrow impurity bands: Application to CaB6_6

Ferromagnetism with high Curie temperature $T_c$, well above room temperature, and very small saturation moment has been reported in various carbon and boron systems. It is argued that the magnetization must be very inhomogeneous with only a small fraction of the sample ferromagnetically ordered. It is shown that a possible source of high $T_c$ within the ferromagnetic regions is itinerant electrons occupying a narrow impurity band. Correlation effects do not reduce the effective interaction which enters the Stoner criterion in the same way as in a bulk band. It is also shown how, in the impurity band case, spin wave excitations may not be effective in lowering $T_c$ below its value given by Stoner theory. These ideas are applied to CaB$_6$ and a thorough review of the experimental situation in this material is given. It is suggested that the intrinsic magnetism of the B$_2$ and O$_2$ dimers might be exploited in suitable structures containing these elements.Comment: 26 pages, 2 figure

Emergence of magnetism in graphene materials and nanostructures

Magnetic materials and nanostructures based on carbon offer unique opportunities for future technological applications such as spintronics. This article reviews graphene-derived systems in which magnetic correlations emerge as a result of reduced dimensions, disorder and other possible scenarios. In particular, zero-dimensional graphene nanofragments, one-dimensional graphene nanoribbons, and defect-induced magnetism in graphene and graphite are covered. Possible physical mechanisms of the emergence of magnetism in these systems are illustrated with the help of computational examples based on simple model Hamiltonians. In addition, this review covers spin transport properties, proposed designs of graphene-based spintronic devices, magnetic ordering at finite temperatures as well as the most recent experimental achievements.Comment: tutorial-style review article -- 18 pages, 19 figure