Expansion of CRISPR Targeting Sites Using an Integrated Gene-Editing System in Apis mellifera

CRISPR/Cas9, a predominant gene-editing tool, has been utilised to dissect the gene function in Apis mellifera. However, only the genomic region containing NGG PAM could be recognised and edited in A. mellifera, seriously hampering the application of CRISPR technology in honeybees. In this study, we carried out the bioinformatics analysis for genome-wide targeting sites of NGG, TTN, and NNGRRT to determine the potential expansion of the SpCas9, SaCas9, Cpf1, and it was found that the targetable spectrum of the CRISPR editing system could be markedly extended via the integrated gene manipulation system. Meanwhile, the single guide RNA (sgRNA)/crRNA of different novel gene editing systems and the corresponding CRISPR proteins were co-injected into honeybee embryos, and their feasibility was tested in A. mellifera. The sequencing data revealed that both SaCas9 and Cpf1 are capable of mediating mutation in A. mellifera, albeit with relatively lower mutagenesis rates for Cpf1 and unstable editing for SaCas9. To our knowledge, our results provide the first demonstration that SaCas9 and Cpf1 can function to induce genome sequence alternation, which extended the editing scope to the targets with TTN and NNGRRT and enabled CRISPR-based genome research in a broader range in A. mellifera.

Some features and problems of application of genomic technologies in the Russian Federation

The article is devoted to the analysis of the experience of regulation and the search for ways to overcome some of the problems that exist in the field of application of genomic technologies. Based on the discovery of differences in the specifics of the choice of mechanisms at the international, regional and national levels in the regulation of relations developing in the field of genomic research, the implementation of their results, approaches are proposed aimed at neutralizing possible negative consequences and resolving existing problems. Special attention is paid to the search for approaches aimed at resolving the problems existing in this area in the Russian Federation. The materials of the article can be useful in both theoretical and practical jurisprudence, and may be of interest for other areas of the human genome research (bioinformatics, medicine etc.).

DNA Isolation: A Method for Improving the Efficiency of DNA Extraction From Clotted Blood Samples

In generally, the genome research of high DNA extraction from clotted blood produced a low quality. The aim of this study is to develop a simple and safe technique to dispering the blood clots by the ball bearing metal shots. The yield and purity of DNA obtained by three steps were significantly different (P < 0.0001), with a higher yield in the modified salting-out method. The salting-out method is simple, efficient and economical for DNA isolation from old clotted blood samples.

GSER (a Genome Size Estimator using R): a pipeline for quality assessment of sequenced genome libraries through genome size estimation

The first step in any genome research after obtaining the read data is to perform a due quality control of the sequenced reads. In a de novo genome assembly project, the second step is to estimate two important features, the genome size and ‘best k -mer’, to start the assembly tests with different de novo assembly software and its parameters. However, the quality control of the sequenced genome libraries as a whole, instead of focusing on the reads only, is frequently overlooked and realized to be important only when the assembly tests did not render the expected results. We have developed GSER, a Genome Size Estimator using R, a pipeline to evaluate the relationship between k -mers and genome size, as a means for quality assessment of the sequenced genome libraries. GSER generates a set of charts that allow the analyst to evaluate the library datasets before starting the assembly. The script which runs the pipeline can be downloaded from http://www.mobilomics.org/GSER/downloads or http://github.com/mobilomics/GSER .

LPMX: A pure rootless composable container system

Delivering tools for genome analysis to users is often difficult given their complex dependencies and conflicts. Container virtualization systems such as Singularity isolate environments, helping developers avoid conflicts between tools. However, they lack composability, an easy way to integrate multiple tools in different containers or multiple tools both in a container and a host, which compromises the use of container systems in genome research. Another issue is that one may not be able to use a single container system of the same version at all sites they use, which discourages the use of container systems. To this end, we present a pure rootless composable container system, LPMX, that provides composability for letting developers easily integrate tools in different existing containers or on host, allowing researchers to compose existing containers. LPMX is pure rootless, so it does not require root privilege neither during installation nor at runtime, allowing researchers to use LPMX across sites without asking permissions from administrators. LPMX provides a pure userspace layered filesystem with at least an order of magnitude lower overhead for launching a new process than existing container systems. LPMX can import Docker and Singularity images.

Comparative Analysis for the Performance of Long-Read-Based Structural Variation Detection Pipelines in Tandem Repeat Regions

There has been growing recognition of the vital links between structural variations (SVs) and diverse diseases. Research suggests that, with much longer DNA fragments and abundant contextual information, long-read technologies have advantages in SV detection even in complex repetitive regions. So far, several pipelines for calling SVs from long-read sequencing data have been proposed and used in human genome research. However, the performance of these pipelines is still lack of deep exploration and adequate comparison. In this study, we comprehensively evaluated the performance of three commonly used long-read SV detection pipelines, namely PBSV, Sniffles and PBHoney, especially the performance on detecting the SVs in tandem repeat regions (TRRs). Evaluated by using a robust benchmark for germline SV detection as the gold standard, we thoroughly estimated the precision, recall and F1 score of insertions and deletions detected by the pipelines. Our results revealed that all these pipelines clearly exhibited better performance outside TRRs than that in TRRs. The F1 scores of Sniffles in and outside TRRs were 0.60 and 0.76, respectively. The performance of PBSV was similar to that of Sniffles, and was generally higher than that of PBHoney. In conclusion, our findings can be benefit for choosing the appropriate pipelines in real practice and are good complementary to the application of long-read sequencing technologies in the research of rare diseases.

Contemporary Challenges and Legal Regulation of Genome Research: Some Considerations

The paper studies some aspects and considerations of the legal regulation of genomic research in the context of modern challenges. Approaches to the formulation of some basic concepts in the field of bioinformatics, such as information, information space, are proposed, the importance of these concepts for legal regulation is substantiated. Approaches to the legal regulation of the commercial use of the results of genomic research in the field of bioinformatics are formulated. Approaches to the legal regulation of the activities of biobanks are proposed based on the analysis of the possibility of using blockchain technology to improve the functioning of the biobank, the possibility of attracting investments through crowdfunding financing. The classification of biobanks according to various criteria is given. Approaches to the formulation of such concepts as donation and parenthood are determined within the framework of the legal regulation of genomic research in the field of human reproduction. The influence of modern challenges associated with the development of science and technology on the formulation of these concepts is considered. Approaches to solving the problem of ensuring a balance of private, group and general interests in the field of legal regulation of genomic research are proposed.