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Recent therapeutic strategies for hemophilia include long-term therapeutic gene expression using adeno-associated virus (AAV) and rebalancing therapy via the downregulation of anticoagulant pathways. However, these approaches have limitations in immune responses or insufficiency to control acute bleeding. Thus, we developed a therapeutic strategy for hemophilia B by a combined rebalancing and human factor 9 (hF9) gene knockin (KI) using a lipid nanoparticle (LNP) and AAV. Antithrombin (AT; Serpin Family C Member 1 [Serpinc1]) was selected as the target anticoagulation pathway for the gene KI. First, the combined use of LNP-clustered regularly interspaced short palindromic repeats (CRISPR) and AAV donor resulted in 20% insertions or deletions (indels) in Serpinc1 and 67% reduction of blood mouse AT concentration. Second, hF9 coding sequences were integrated into approximately 3% of the target locus. hF9 KI yielded approximately 1,000 ng/mL human factor IX (hFIX) and restored coagulation activity to a normal level. LNP-CRISPR injection caused sustained AT downregulation and hFIX production up to 63 weeks. AT inhibition and hFIX protein-production ability could be maintained by the proliferation of genetically edited hepatocytes in the case of partial hepatectomy. The co-administration of AAV and LNP showed no severe side effects except random integrations. Our results demonstrate hemophilia B therapy by a combination of rebalancing and hF9 KI using LNP and AAV.

Pseudotyped virus systems that incorporate viral proteins have been widely employed for the rapid determination of the effectiveness and neutralizing activity of drug and vaccine candidates in biosafety level 2 facilities. We report an efficient method for producing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pseudovirus with dual luciferase and fluorescent protein reporters. Moreover, using the established method, we also aimed to investigate whether Korean Red Ginseng (KRG), a valuable Korean herbal medicine, can attenuate infectivity of the pseudotyped virus.A pseudovirus of SARS-CoV-2 (SARS-2pv) was constructed and efficiently produced using lentivirus vector systems available in the public domain by the introduction of critical mutations in the cytoplasmic tail of the spike protein. KRG extract was dose-dependently treated to Calu-3 cells during SARS2-pv treatment to evaluate the protective activity against SARS-CoV-2.The use of Calu-3 cells or the expression of angiotensin-converting enzyme 2 (ACE2) in HEK293T cells enabled SARS-2pv infection of host cells. Coexpression of transmembrane protease serine subtype 2 (TMPRSS2), which is the activator of spike protein, with ACE2 dramatically elevated luciferase activity, confirming the importance of the TMPRSS2-mediated pathway during SARS-CoV-2 entry. Our pseudovirus assay also revealed that KRG elicited resistance to SARS-CoV-2 infection in lung cells, suggesting its beneficial health effect.The method demonstrated the production of SARS-2pv for the analysis of vaccine or drug candidates. When KRG was assessed by the method, it protected host cells from coronavirus infection. Further studies will be followed for demonstrating this potential benefit.

Chemical bath-deposited SnO2 widely used as an electron transport layer has hydroxyl defect, which is responsible for degradation of perovskite solar cell. Surface passivation of SnO2 with 4-fluorothiophenol improves stability.

Quasi-two-dimensional (Quasi-2D) van der Waals (vdW) materials can be mechanically or chemically exfoliated down to monolayer because of their strong intralayer bonding and the weak interlayer vdW interaction. Thanks to this unique property, one can often find exotic thickness-dependent electronic properties from these quasi-2D vdW materials, which can lead to band gap opening, emerging superconductivity, or enhanced charge density waves with decreasing thickness. Surface-sensitive scanning tunneling microscopy (STM) can provide direct observation of structural and electronic characteristics of such layered materials with atomic precision in real space. However, it is very challenging to preserve the intrinsic surfaces of air-sensitive quasi-2D materials between preparation and measurement. In addition, vdW 2D crystals after exfoliation are extremely hard to explore with a typical STM setup due to their small size (< 10 {\mu}m). Here, we present a straightforward method compatible with any STM setup having optical access: (1) exfoliating and/or stacking layered materials in a glove box, (2) transferring them to an ultra-high vacuum STM chamber using a suitcase without exposure to air, and (3) navigating surface to locate exfoliated vdW 2D flakes with different thicknesses. We successfully demonstrated that the clean surfaces of the air-sensitive Fe$_3$GeTe$_2$ can be effectively protected from unwanted oxidation during transfer. Furthermore, our method provides a simple but useful way to access a specific tiny stack of layered materials without any ex-situ fabrication processes for STM navigation. Our experimental improvement will open up a new way to investigate air-sensitive layered vdW materials with various thicknesses via surface-sensitive techniques including STM., Comment: 5 pages, 5 figures

Funder: GIST Research Institute, Conducting polymers are a key component for developing wearable organic electronics, but tracking their redox processes at the nanoscale to understand their doping mechanism remains challenging. Here we present an in-situ spectro-electrochemical technique to observe redox dynamics of conductive polymers in an extremely localized volume (

We present the results of a radio multi-frequency ($\rm 3-340~GHz$) study of the blazar 3C~454.3. After subtracting the quiescent spectrum corresponding to optically thin emission, we found two individual synchrotron self-absorption (SSA) features in the wide-band spectrum. The one SSA had a relatively low turnover frequency ($\nu_{\rm m}$) in the range of $\rm 3-37~GHz$ (lower $\nu_{\rm m}$ SSA spectrum, LSS), and the other one had a relatively high $\nu_{\rm m}$ of $\rm 55-124~GHz$ (higher $\nu_{\rm m}$ SSA spectrum, HSS). Using the SSA parameters, we estimated magnetic field strengths at the surface where optical depth $\tau=1$. The estimated magnetic field strengths were $\rm >7~mG$ and $\rm >0.2~mG$ for the LSS and HSS, respectively. The LSS emitting region was magnetically dominated before the June 2014 $\gamma$-ray flare. The quasi-stationary component (C), $\sim 0.6~{\rm mas}$ apart from the 43 GHz radio core, became brighter than the core with decreasing observing frequency, and we found that component C was related to the LSS. A decrease in jet width was found near component C. As a moving component, K14 approached component C, and the flux density of the component was enhanced while the angular size decreased. The high intrinsic brightness temperature in the fluid frame was obtained as $T_{\rm B, int} \approx (7.0\pm1.0) \times 10^{11}~{\rm K}$ from the jet component after the 2015 August $\gamma$-ray flare, suggesting that component C is a high-energy emitting region. The observed local minimum of jet width and re-brightening behavior suggest a possible recollimation shock in component C., Comment: 16 pages, 12 figures, 5 tables, Accepted for publication in MNRAS

Flexible micro-light-emitting diodes (f-μLEDs) have been regarded as an attractive light source for the next-generation human-machine interfaces, thanks to their noticeable optoelectronic performances. However, when it comes to their practical utilizations fulfilling industrial standards, there have been unsolved reliability and durability issues of the f-μLEDs, despite previous developments in the high-performance f-μLEDs for various applications. Herein, highly robust flexible μLEDs (f-HμLEDs) with 20 × 20 arrays, which are realized by a siloxane-based organic-inorganic hybrid material (SHM), are reported. The f-HμLEDs are created by combining the f-μLED fabrication process with SHM synthesis procedures (i.e., sol-gel reaction and successive photocuring). The outstanding mechanical, thermal, and environmental stabilities of our f-HμLEDs are confirmed by a host of experimental and theoretical examinations, including a bending fatigue test (10

Supplementary Table 1: List of primers used for quantitative real time PCR. Supplementary Table. 2: List of primers used for construction of pGL2 and pMIR reporter plasmids. Supplementary Table. 3: List of primers used to generate PCR amplicons for pyrosequencing analysis. Supplementary Table. 4: Sequencing primers used for pyrosequencing. Supplementary Table. 5: Primers used for qPCR analysis in ChIP assay.

A subset of primary breast tumors expresses a 308-gene signature that correlated with gene signatures specifically expressed in immune cells.