Radon and magnetic anomalies in the territory of the city Kyiv: environmental aspect

Magnetic and radon anomalous fields are essential geophysical entities of environment which determine to a considerable degree its ecological state. Revelation of magnetic and radon anomalous zones is an urgent task for the studies of geo-environmental state of the territory and appraisal of its ecological safety. The greatest danger is produced by the areas where the faults are localized with radon anomalies related in the upper part of the Earth crust. For the territory of Kyiv geological characteristics of the studied area has been reviewd, schemes of regional and accompanying faults have been adjusted and completed, maps of module and anomalies of module of geomagnetic field induction as well as a scheme of radon anomalies in water and subsoil air have been elaborated. Connection between radon anomalies with faults and natural magnetic fields has been shown which consisted in correlation of radon anomalies with negative and small positive values of anomalous magnetic field and related to thick zones of fracturing with active fluidal-dynamic activity and the faults of diagonal and orthogonal systems being the zones of radon emanation into atmosphere. Radon anomalies in subsoil air are mainly controlled by Irpin-Borshchagivka, Pushcha-Vodytsya, Kyiv and Darnytsya faults of northwestern-northern strike and Petrivka-Pukhivka and Glevakha-Brovary faults of northwestern strike. For some stations of Kyiv metro the excess of environmental norms of magnetic field and radon anomalies has been determined which can be explained by constructive features of metro compartments and their location in fault zones. It has been shown that radon anomalies are spatially and temporally stabile and magnetic anomalies, especially in metro, are characterized by considerable changes of magnetic field in both low-frequency and high-frequency spectra of their variation. Combined analysis of the fault tectonic scheme, magnetic and radon anomalies will permit to find out environmentally hazardous zones in the territory of Kyiv and out of it more reasonably.

Orthogonal CRISPR-Cas genome editing and efficient inhibition with anti-CRISPRs in zebrafish embryos

The CRISPR-Cas universe continues to expand. The type II CRISPR-Cas system from Streptococcus pyogenes (SpCas9) is most widely used for genome editing due to its high efficiency in cells and organisms. However, concentrating on a single CRISPR-Cas system limits options for multiplexed editing. We hypothesized that CRISPR-Cas systems originating from different bacterial species could operate simultaneously and independently due to their distinct single-guide RNAs (sgRNAs) or CRISPR-RNAs (crRNAs), and protospacer adjacent motifs (PAMs). Additionally, we hypothesized that CRISPR-Cas activity in zebrafish could be regulated through the expression of inhibitory anti-CRISPR (Acr) proteins. Here, we use a simple mutagenesis approach to demonstrate that CRISPR-Cas systems from Streptococcus pyogenes (SpCas9), Streptococcus aureus (SaCas9), and Lachnospiraceae bacterium (LbCas12a, previously known as LbCpf1) are highly effective, orthogonal systems capable of operating simultaneously in zebrafish. We also demonstrate that type II Acrs are effective inhibitors of SpCas9 in zebrafish. These results indicate that at least three orthogonal CRISPR-Cas systems and two anti-CRISPR proteins are functional in zebrafish embryos. These CRISPR-Cas systems and Acr proteins will enable combinatorial and intersectional strategies for spatiotemporal control of genome editing in zebrafish.