Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping

Prolonged expression of the CRISPR-Cas9 nuclease and gRNA from viral vectors may cause off-target mutagenesis and immunogenicity. Thus, a transient delivery system is needed for therapeutic genome editing applications. Here, we develop an extracellular nanovesicle-based ribonucleoprotein delivery system named NanoMEDIC by utilizing two distinct homing mechanisms. Chemical induced dimerization recruits Cas9 protein into extracellular nanovesicles, and then a viral RNA packaging signal and two self-cleaving riboswitches tether and release sgRNA into nanovesicles. We demonstrate efficient genome editing in various hard-to-transfect cell types, including human induced pluripotent stem (iPS) cells, neurons, and myoblasts. NanoMEDIC also achieves over 90% exon skipping efficiencies in skeletal muscle cells derived from Duchenne muscular dystrophy (DMD) patient iPS cells. Finally, single intramuscular injection of NanoMEDIC induces permanent genomic exon skipping in a luciferase reporter mouse and in mdx mice, indicating its utility for in vivo genome editing therapy of DMD and beyond

ハリエンジュにおけるチロース発生の季節とそれに伴う柔細胞構造の変化

ハリエンジュの樹幹から辺材部を定期的にサンプリングした。この光顕および走査電顕観察から, 9月下旬から10月下旬にかけて当年輪の孔圏道管に, それに接した軸方向柔細胞からチロースが多く発生することが明らかとなった。つぎに, この軸方向柔細胞をはじめ, チロースおよび道管周辺の各種細胞の構造を透過電顕で調べた。チロース芽発生前の9月には細胞は休止型あるいは貯蔵型の構造を示し living fiber も starch grains と lipid droplets を保有していた。チロース芽が活発に発生している10月になると, 道管に接した軸方向柔細胞とチロース芽にゴルジ体, r-ER, ミトコンドリア, リボゾーム等が充満しており, 典型的な活動型の細胞構造となった。道管がチロースで閉そくされた12月には, 道管に接した軸方向柔細胞ではゴルジ体, r-ERは減少しており, 大径の脂質粒が現われた。また液胞中には代謝副産物と思われる顆粒が発生していた

Identification of Factors Necessary for Enabling Technology-Based Dietary Record Surveys: A Qualitative Focus Group Interview with Japanese Dietitians

Weighed food records together with an in-person interview approach constitute the most basic methods used to estimate energy and nutrient intakes in dietary surveys. In the background of the coronavirus disease-2019 pandemic, the need for non-face-to-face dietary surveys using information and communication technology (ICT) is increasing. We aimed to evaluate ICT-based dietary record surveys and identify factors that may enable this survey method to become more widely used in the future. We conducted a non-face-to-face survey of dietary records of 44 Japanese individuals, maintained by dietitians using dietary photography and video conferencing services. We conducted a focus group interview with the six dietitians who conducted that survey. Their opinions on the factors necessary to popularize ICT-based dietary survey method were analyzed. In the focus group interview, dietitians highlighted fewer restrictions on time and place as positive aspects. Negative aspects included insufficient skills to operate computers, difficulty in hearing, and understanding facial expressions using ICT. We identified three main factors for enabling widespread use of ICT-based dietary record survey: individual skill, device and technology, and social environmental factors. This suggests that a comprehensive approach is necessary for popularizing the use of ICT in dietary surveys

Chemical evidence for the origin of the cold water belt along the northeastern coast of Hokkaido

In the southwestern Okhotsk Sea, the cold water belt (CWB) is frequently observed on satellite images offshore of the Soya Warm Current flowing along the northeastern coast of Hokkaido, Japan, during summertime. It has been speculated that the CWB is upwelling cold water that originates from either subsurface water of the Japan Sea off Sakhalin or bottom water of the Okhotsk Sea. Hydrographic and chemical observations (nutrients, humic-type fluorescence intensity, and iron) were conducted in the northern Japan Sea and southwestern Okhotsk Sea in early summer 2011 to clarify the origin of the CWB. Temperature-salinity relationships, vertical distributions of chemical components, profiles of chemical components against density, and the (NO3 + NO2)/PO4 relationship confirm that water in the CWB predominantly originates from Japan Sea subsurface water