Search

The bacteria-derived clustered regularly interspaced short palindromic repeat (CRISPR)–Cas systems are powerful tools for genome engineering. Recently, in addition to Cas protein engineering, the improvement of guide RNAs are also performed, contributing to broadening the research area of CRISPR–Cas9 systems. Here we develop a fusion guide RNA (fgRNA) that functions with both Cas9 and Cpf1 proteins to induce mutations in human cells. Furthermore, we demonstrate that fgRNAs can be used in multiplex genome editing and orthogonal genome manipulation with two types of Cas proteins. Our results show that fgRNAs can be used as a tool for performing multiple gene manipulations., Cas9 and Cpf1 have both been adapted for genome engineering, editing and gene expression regulation. Here the authors design a fusion guide RNA that can interact with both proteins for multiple and orthogonal genome manipulation.

Glass-ceramics were developed many years ago and have been applied in many fields such as electronics, chemistry, optics, etc. Much is already known about glass-ceramic technology, but many challenges in glass-ceramic research are still unresolved. Recently, large amounts of slag have steadily increased in the steel industry as by-products. To promote recycling of industrial waste, including steel industry slags, many studies have been performed on the fabrication of basalt-based high-strength glass-ceramics. In this study, we have fabricated such ceramics using various slags to replace high performance cast-basalt, which is currently imported. Glass-ceramic material was prepared in similar chemical compositions with commercial cast-basalt through a pyro process using slags and power plant by-product (Fe-Ni slag, converter slag, dephosphorization slag, Fly ash). The properties of the glass-ceramic material were characterized using DTA, XRD, and FE-SEM; measurements of compressive strength, Vicker`s hardness, and abrasion were carefully performed. It is found that the prepared glass-ceramic material showed better performance than that of commercial cast-basalt.

The originally published version of this Article contained an error in the spelling of the author Da-eun Kim, which was incorrectly given as Da-Eun Kim. Furthermore, in Figure 1a, the Cas9 protein was positioned incorrectly during typesetting. These errors have now been corrected in both the PDF and HTML versions of the Article.

Genome editing using programmable nucleases such as CRISPR/Cas9 or Cpf1 has emerged as powerful tools for gene knock-out or knock-in in various organisms. While most genetic diseases are caused by point mutations, these genome-editing approaches are inefficient in inducing single-nucleotide substitutions. Recently, Cas9-linked cytidine deaminases, named base editors (BEs), have been shown to convert cytidine to uridine efficiently, leading to targeted single-base pair substitutions in human cells and organisms. Here, we first report on the generation of Xenopus laevis mutants with targeted single-base pair substitutions using this RNA-guided programmable deaminase. Injection of base editor 3 (BE3) ribonucleoprotein targeting the tyrosinase (tyr) gene in early embryos can induce site-specific base conversions with the rates of up to 20.5%, resulting in oculocutaneous albinism phenotypes without off-target mutations. We further test this base-editing system by targeting the tp53 gene with the result that the expected single-base pair substitutions are observed at the target site. Collectively, these data establish that the programmable deaminases are efficient tools for creating targeted point mutations for human disease modeling in Xenopus.

The present meta-analysis examined the stability of vocational interests from early adolescence (age 12) to middle adulthood (age 40). Stability was represented by rank-order and profile correlations. Interest stability remained unchanged during much of adolescence and increased dramatically during the college years (age 18-21.9), where it remained for the next 2 decades. Analyses of potential moderators showed that retest time interval was negatively related to interest stability and that rank-order stability was less stable than profile stability. Although cohort standings did not moderate stability, interests of the 1940s birth cohort were less stable than those of other cohorts. Furthermore, interests reflecting hands-on physical activities and self-expressive/artistic activities were more stable than scientific, social, enterprising, and clerical interests. Vocational interests showed substantial continuity over time, as evidenced by their higher longitudinal stability when compared with rank-order stability of personality traits. The findings are discussed in the context of psychosocial development.

Peroxisome proliferator-activated receptor alpha and estrogen are believed to be involved in metabolic changes leading to obesity. To test this relationship, we divided female wildtype and PPAR alpha-deficient mice fed on a high fat diet into the following groups: mock-operated, ovariectomized (OVX), and E(2)-treated. The visceral white adipose tissue and plasma cholesterol levels were increased significantly in wild type OVX and decreased in the E(2)-treated group, but interestingly not in PPAR alpha-deficient mice. The mRNA levels of lipoprotein lipase in adipose tissue were also increased in only wild type OVX and decreased significantly in E(2)-treated mice. These novel results suggest the possibility of signaling crosstalk between PPAR alpha and E2, causing obesity in vivo.

Considerable controversy exists in determining the role of peroxisome proliferator-activated receptor-alpha PPARalpha) on obesity. Previous reports demonstrated that PPARalpha is a critical modulator of lipid homeostasis, but the overt, obese phenotypic characterization in the strain of PPAR deficient (PPARalpha-/-) mice is influenced by other factors, including diet and genetics. Therefore, it is necessary to establish the phenotypic characterization of PPARalpha-/- mice prior to the obesity-related study. In this study, we observed phenotype of PPARalpha-/- mice on mixed genetic background (C57BL/6Nx129/Sv) fed a high fat diet for 16 weeks. PPARalpha-/- mice, regardless of sex, raised body growth rate significantly comparing with wild type and showed male-specific fatty change in the liver. They were shown to lack hepatic induction of PPARalpha target genes encoding enzymes for fatty acid beta-oxidation.