CRISPR/Cas9, a universal tool for genomic engineering

The CRISPR/Cas9 system was initially described as an element of archeal and bacterial immunity, but gained much attention recently for its outstanding ability to be programmed to target any genomic loci through a short 20-nucleotide sgRNA region. Here we review some modern applications of the CRISPR/Cas9 system. First, we describe the basic mechanism of the CRISPR/Cas9 DNA recognition and binding, focusing in particular on its off-target activity. The CRISPR/Cas9 off-target activity refers to a non-specific recognition of genomic sites that have partial homology with sgRNA, occasionally resulting in unwanted mutations throughout the genome. We also note some recent improvements for enhancing Cas9 specificity or adding new functions to the system. Since Cas9-related hype is mostly driven by its remarkable potential for gene therapy and genome engineering, the latest CRISPR/Cas9 applications in these areas are also covered in our review. For instance, the CRISPR/Cas9 was recently used to control HIV infection and to repair genetic abnormalities, such as Duchenne muscular dystrophy or retinitis pigmentosa, both in cell cultures and rodent models. A programmable nature of CRISPR/Cas9 facilitates the creation of transgenic organisms through sitespecific gene mutations, knock-ins or large chromosomal rearrangements (deletions, inversions and duplications). CRISPR/Cas9 proved to be especially useful in pronuclear microinjections of farm animals as well, having strong impact on biotechnology. In addition, we review Cas9-augmented genetic screens that allow an unbiased search for new genes and pathways involved in a plethora of biological aspects, owing to Cas9 efficiency and versatility. Finally, we argue that gene drivers based on CRISPR/ Cas9 represent a powerful tool to modify ecosystems in the nearest future

Legal certainty of legality principle in the legislation of the continental and anglo-american system

In this article, through the comparative legal analysis of domestic, as well as foreign constitutional and criminal law norm content, various approaches to the principle of legality and its main features were examine

Genome editing using CRISPR/ Cas9 system: a practical guide

Over the past few years, the CRISPR/Cas techniques have become a revolution in genome editing. Since the original paper on CRIPSR/Cas9 genome editing, researches have proposed numerous modifications of the key components of the CRISPR/Cas9 system to make it extremely efficient. Nowadays, CRISPR/Cas systems can be used not only to modify genomes, but also to control expression levels of defined genes, visualize loci of interest in the space of living cell nuclei, change methylation status of mammalian CpG sites, and to serve many other purposes. Due to an extremely high efficacy and ease of usage, the CRISPR/ Cas system has been employed in a large number of studies in various areas of biology and biotechnology. We have recently published a review describing various CRISPR/Cas systems, mechanisms of their functioning, and applications of the techniques in details. Despite the broad range of potential applications of CRISPR/Cas systems, they are mostly used for genome editing. And, however simple the system may be, there is a number of potential pitfalls on the way towards its use in CRISPR/Cas- naïve laboratory settings. In this article, we describe protocols of CRISPR/Cas9 system generation. We start with a short description of theoretical aspects underlying Cas9-mediated genome editing. Next, we describe a step-by-step protocol of guide RNA vector design and assembly, and several ways of qualitative and quantitative evaluations of the system. Finally, we report protocols of genome editing for modification of embryonic stem cells and zygotes

International standards for the safety of persons assisting in criminal justice

The objective ofthe research was to analyze some international standards for the safety of people who attend criminal justice from different approaches and perspectives of analysis. Based on a meaningful analysis of the provisions of international and regional regulatory legal acts, the document presents approaches to the formation of standards to ensure the safety of persons who contribute to criminal justic

Low-frequency noise in nFinFETs of different dimensions processed in strained and non-strained SOI wafers

The results of low-frequency noise investigation in fully-depleted (FD) nFinFETs of Weff = 0.02 to 9.87 µm, Leff = 0.06 to 9.9 µm, processed on standard (SOI) and strained (sSOI) wafers are presented. It is shown that the McWhorter noise is typical at zero back gate voltage for the devices studied and the density of the corresponding noisy traps in the SiO₂ portion of the gate oxide is, as a rule, much higher than that in the HfO2 portion. The results on the McWhorter noise are used for studying the behavior of the electron mobility µ and the free electron density NS in the channel at V* ≥ 0.4 V where V* is the gate overdrive voltage. It is also shown that the Linear Kink Effect (LKE) Lorentzians appear in the low-frequency noise spectra at an accumulation back gate voltage and that the parameters of those Lorentzians are different for the sSOI and SOI nFinFETs. This is the first observation of the LKE noise under a back-gate accumulation bias for sufficiently wide nMuGFET

EFFECT OF LEAD ON STRUCTURAL CHANGES IN LIVER OF WISTAR RATS UNDER THE CONDITIONS OF ACUTE EXPERIMENT

The purpose of this study was to evaluate the structural transformations of Wistar rats liver after oral administration of lead salts. Under the conditions of acute experiment during 5 days the Wistar rats were orally administered the lead acetate solution in the amount of 3 mg/kg. Histological studies were carried out at OSMU upon the completion of the experiment. Effect of toxic doses of lead causes symptoms of both steatosis and hydropic degeneration of the liver. It is assumed that the appearance of hepatic steatosis reflects the reaction of hepatocytes to hemic hypoxia caused by the action of lead, while the signs of hydropic degeneration expressed in varying degrees reflect its direct toxic effect

Legal certainty of legality principle in the legislation of the continental and anglo-american system

In this article, through the comparative legal analysis of domestic, as well as foreign constitutional and criminal law norm content, various approaches to the principle of legality and its main features were examine

Anopheles mosquitoes reveal new principles of 3D genome organization in insects.

Chromosomes are hierarchically folded within cell nuclei into territories, domains and subdomains, but the functional importance and evolutionary dynamics of these hierarchies are poorly defined. Here, we comprehensively profile genome organizations of five Anopheles mosquito species and show how different levels of chromatin architecture influence each other. Patterns observed on Hi-C maps are associated with known cytological structures, epigenetic profiles, and gene expression levels. Evolutionary analysis reveals conservation of chromatin architecture within synteny blocks for tens of millions of years and enrichment of synteny breakpoints in regions with increased genomic insulation. However, in-depth analysis shows a confounding effect of gene density on both insulation and distribution of synteny breakpoints, suggesting limited causal relationship between breakpoints and regions with increased genomic insulation. At the level of individual loci, we identify specific, extremely long-ranged looping interactions, conserved for ~100 million years. We demonstrate that the mechanisms underlying these looping contacts differ from previously described Polycomb-dependent interactions and clustering of active chromatin