A bibliometric analysis-based literature review of the relationship between sustainable water management and green innovations in the agricultural sector.

Global water demand has grown intensively over the last three decades, and the predictions suggest this trend will continue. Sustainable Water Management (SWM) defines water-based principles and action frameworks interconnecting societal, economic, and environmental aspects to establish and maintain good practices serving long-term objectives related to water resources. Water scarcity, deterioration of water quality, less effective water technologies, hydrological changes caused by climate change, and increased water demand require the thorough revision of conventional approaches, new methods, and new policy measures. The research methodology in this paper includes a comprehensive review and bibliometric analysis of relevant literature on water management and sustainable development, including empirical studies, theoretical frameworks, and policy documents. The study explores the conceptual context of SWM, reveals the barriers hindering its core progress, evaluates the impact of green innovations on the development of novel operations, and gets an insight into the current policy and regulatory framework for SWM. Besides giving a review of the current practices and perspectives in SWM, the results of this study contribute to a deeper understanding of the complex relationship between sustainable water management and green innovations in the agricultural sector and provide possible directions toward adopting effective strategies and policies to promote a more intense permeation of the SWM approach.

A convenient method to pre-screen candidate guide RNAs for CRISPR/Cas9 gene editing by NHEJ-mediated integration of a 'self-cleaving' GFP-expression plasmid.

The efficacies of guide RNAs (gRNAs), the short RNA molecules that bind to and determine the sequence specificity of the Streptococcus pyogenes Cas9 nuclease, to mediate DNA cleavage vary dramatically. Thus, the selection of appropriate target sites, and hence spacer sequence, is critical for most applications. Here, we describe a simple, unparalleled method for experimentally pre-testing the efficiencies of various gRNAs targeting a gene. The method explores NHEJ-cloning, genomic integration of a GFP-expressing plasmid without homologous arms and linearized in-cell. The use of 'self-cleaving' GFP-plasmids containing universal gRNAs and corresponding targets alleviates cloning burdens when this method is applied. These universal gRNAs mediate efficient plasmid cleavage and are designed to avoid genomic targets in several model species. The method combines the advantages of the straightforward FACS detection provided by applying fluorescent reporter systems and of the PCR-based approaches being capable of testing targets in their genomic context, without necessitating any extra cloning steps. Additionally, we show that NHEJ-cloning can also be used in mammalian cells for targeted integration of donor plasmids up to 10 kb in size, with up to 30% efficiency, without any selection or enrichment.