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The nuclease-dead Cas9 (dCas9) has been reprogrammed for transcriptional activation by fusing dCas9 to a transcriptional activation domain. In the presence of a guide RNA (gRNA), the dCas9 fusions specifically bind to regions of a promoter to activate transcription. Significant amount of effort has been directed toward the identification and optimization of the fusions of dCas9-activation domain, but very little is known about the impact of gRNA target positions within a promoter in plants on transcriptional activation efficiency. The dCas9-6TAL-VP128 system (dCas9-TV) has been optimized to activate transcription in plants. Here we use the dCas9-TV to activate transcription of OsWOX11 and OsYUC1, two genes that cause dramatic developmental phenotypes when overexpressed. We designed a series of gRNAs targeting the promoters of the two genes. We show that gRNAs that target regions within 350 bp upstream of the transcription start site were most effective in transcriptional activation. Moreover, we show that using two gRNAs that simultaneously target two discrete sites in a promoter can further enhance transcription. This work provides guidelines for designed transcriptional activation through CRISPR/dCas9 systems.

The tailoring of the surface properties of cellulose nanocrystals (CNCs) to meet various requirements in environmental, food, and material areas has always been of great interest. In this study, the surface chemistry of CNCs was noncovalently modified by cetyltrimethylammonium bromide (CTAB), followed by characterizations and an investigation into its application as a coating material for interfacial interaction over various substrates. Due to the CTAB modification, the surface charge of the CNCs was neutralized, resulting in an increased size of each nanocrystal at the aqueous status and the aggregated microfibers when dried up. The CTAB modification not only decreased the crystallinity of the samples from 48.57% to 9.12%, but also reasonably hydrophobized the CNCs and decreased their total surface energy. Finally, the adsorption behavior of the CNCs and CTAB-CNCs over nonionic, anionic, and cationic polymers was investigated by ellipsometry. Based on the thickness of the CNC and CTAB-CNC layers over 2-Hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC), and polyethyleneimine (PEI), we proposed that the adsorption behavior was overall influenced by electrostatic interaction, hydrogen bonding, and van der Waals forces, and the thickness of the adsorbed layers could be impacted by both the surface charge and the size of the crystals. [ABSTRACT FROM AUTHOR]

Pyrolysis and gasification are considered as a means of producing renewable energy and improving energy sustainability, which has become attractive renewable technologies to many countries. Unlike other studies that are conducted in small scale, this study aims to aggregate the economic and environmental effects such as agricultural benefits, energy sale, and carbon sequestration to provide more detailed information to decision-makers before these projects are widely employed. This study first employs a lifecycle assessment to investigate the feasibility, profitability, and emission reduction of four major pyrolysis and gasification technologies using crop residuals, and then conducts a sensitive analysis to examine the most influential factors. The results indicate that the intermediate pyrolysis with rice straw and slow pyrolysis from corn stover could offset the carbon dioxide the most. However, the pyrolysis value is also sensitive to production of the feedstock used. Value adding of stover-based biochar under fast pyrolysis improves profitability but other technologies do not have such patterns. Additionally, while gasification can generate considerable amount of renewable electricity, it yields almost zero percent of biochar that can be used as a soil amendment, and thus its contribution to agricultural sector is trivial. [ABSTRACT FROM AUTHOR]

Among the 13 T1 plants analyzed, eight of them were quadruple mutants (Figure 1g), (ii) The gene-editing efficiency differed significantly among the four target genes (Figure 1g), (iii) I GENE1 i , I GENE2, i and I GENE3 i were not null in nine T1 plants of the TKC-M1 because they contained deletions of 3 or 9 bp. Editing gene families by CRISPR/Cas9: accelerating the isolation of multiple transgene-free null mutant combinations with much reduced labor-intensive analysis Keywords: CRISPR; transgene-free; multi; TKC; rice EN CRISPR transgene-free multi TKC rice 241 243 3 01/17/22 20220201 NES 220201 Genetically elucidating functions of gene families often requires simultaneous disruption of several family members to overcome redundancy. [Extracted from the article]