Direct Injection of CRISPR/Cas9-Related mRNA into Cytoplasm of Parthenogenetically Activated Porcine Oocytes Causes Frequent Mosaicism for Indel Mutations

Some reports demonstrated successful genome editing in pigs by one-step zygote microinjection of mRNA of CRISPR/Cas9-related components. Given the relatively long gestation periods and the high cost of housing, the establishment of a single blastocyst-based assay for rapid optimization of the above system is required. As a proof-of-concept, we attempted to disrupt a gene (GGTA1) encoding the α-1,3-galactosyltransferase that synthesizes the α-Gal epitope using parthenogenetically activated porcine oocytes. The lack of α-Gal epitope expression can be monitored by staining with fluorescently labeled isolectin BS-I-B₄ (IB4), which binds specifically to the α-Gal epitope. When oocytes were injected with guide RNA specific to GGTA1 together with enhanced green fluorescent protein (EGFP) and human Cas9 mRNAs, 65% (24/37) of the developing blastocysts exhibited green fluorescence, although almost all (96%, 23/24) showed a mosaic fluorescent pattern. Staining with IB4 revealed that the green fluorescent area often had a reduced binding activity to IB4. Of the 16 samples tested, six (five fluorescent and one non-fluorescent blastocysts) had indel mutations, suggesting a correlation between EGFP expression and mutation induction. Furthermore, it is suggested that zygote microinjection of mRNAs might lead to the production of piglets with cells harboring various mutation types.     

Hopping transport in the space-charge region of p-n structures with InGaN/GaN QWs as a source of excess 1/f noise and efficiency droop in LEDs

Semiconductors - It is shown that the emission efficiency and the 1/f noise level in light-emitting diodes with InGaN/GaN quantum wells correlate with how the differential resistance of a diode...     

Collective reflection strategy for moderating conformity tendency and promoting reflective judgment performance

Reflective judgement is crucial for medical-related practitioners in dealing with controversial issues. However, the conformity phenomenon is likely to occur and interfere with reflective judgement learning during interactive activities. Effective strategies are required to moderate the conformity behaviour tendency (CBT) and improve reflective judgement performance (RJP). This study demonstrates two significant results: (a) Compared with the guided self-reflection learning strategy, the online collective reflection (OCR) learning strategy effectively weakened the learners' general CBT while dealing with professional controversial issues; and (b) a significantly negative correlation between the RJP achieved and the change of CBT in online environment was detected in the OCR group. The implications and potential applications in higher education were discussed. Further studies are needed to confirm the long-term effects and the extending application to other professional studies.     

Triphenylphosphonium (TPP)-Based Antioxidants: A New Perspective on Antioxidant Design

Mitochondrial oxidative damage and dysfunction contribute to a wide range of human diseases. Considering the limitation of conventional antioxidants and that mitochondria are the main source of reactive oxygen species (ROS) which induce oxidative damage, mitochondria-targeted antioxidants which can selectively block mitochondrial oxidative damage and prevent various types of cell death have been widely developed. As a lipophilic cation, triphenylphosphonium (TPP) has been commonly used in designing mitochondria-targeted antioxidants. Conjugated with the TPP moiety, antioxidants can achieve more than 1000-fold higher mitochondrial concentration depending on cell membrane potentials and mitochondrial membrane potentials. Herein we discuss the deficiencies of conventional antioxidants and the advantages of mitochondrial targeting, and review various types of TPP-based mitochondria-targeted antioxidants. These provide theoretical and background support for the design of new anti-oxidant.