The targeted degradation of membrane proteins would afford an attractive and general strategy for treating various diseases that remain difficult with the current proteolysis-targeting chimera (PROTAC) methodology. We herein report a covalent nanobody-based PROTAC strategy, termed GlueTAC, for targeted membrane protein degradation with high specificity and efficiency. We first established a mass-spectrometry-based screening platform for the rapid development of a covalent nanobody (GlueBody) that allowed proximity-enabled cross-linking with surface antigens on cancer cells. By conjugation with a cell-penetrating peptide and a lysosomal-sorting sequence, the resulting GlueTAC chimera triggered the internalization and degradation of programmed death-ligand 1 (PD-L1), which provides a new avenue to target and degrade cell-surface proteins.

Objectives: We aimed to (1) assess parental hesitancy about category A (Expanded Program on Immunization (EPI)) and B (non-EPI) vaccines, (2) assess parental willingness for COVID-19 and influenza vaccinations, and (3) explore the association of vaccination hesitancy of parents and healthcare workers (HCWs). Methods: The study was performed in Wuxi, eastern China between 21 September 2020 and 17 October 2020. Parents of children aged <18 years and HCWs were recruited from the selected immunization clinics. Vaccine hesitancy was assessed using the Strategic Advisory Group of Experts (SAGE) vaccine hesitancy survey (VHS) by summing the total score for 10 items (maximum 50 points). Results: A total of 3009 parents and 86 HCWs were included in the analysis. The category A VHS scores were significantly higher than the category B VHS scores (p = 0.000). Overall, 59.3% and 52.4% of parents reported willingness to avail COVID-19 and influenza vaccination for their children, respectively; 51.2% of the HCWs wanted to be vaccinated against COVID-19. Parental category B VHS scores were associated with HCW category B VHS scores (r = 0.928, p = 0.008). Conclusions: In China, parents are more hesitant about category B vaccines than category A vaccines. More than 40% of parents showed hesitancy and a refusal to use COVID-19 and influenza vaccines.

SummaryTapetum development and pollen production are regulated by a complex transcriptional network that consists of a group of tapetum-specific Arabidopsis transcription factors (TFs). Among these TFs, DEFECTIVE IN TAPETAL DEVELOPMENT AND FUNCTION 1 (TDF1) encodes an R2R3 MYB factor, and ABORTED MICROSPORE (AMS) encodes a basic helixloop-helix (bHLH) factor. However, knowledge regarding the regulatory role of TDF1 in anther development remains limited.Here, we discovered that TDF1 directly regulates AMS via an AACCT cis-element. We found the precocious AMS transcript and absence of AMS protein in ams À/À gpTDF1:AMS-FLAG lines, suggesting the timing of the TDF1-regulated AMS expression is a prerequisite for AMS functioning. We found that TDF1 interacts with AMS. Additionally, the TDF1-AMS complex additively promotes the expression of AMS-regulated genes, suggesting that TDF1 and AMS regulate the downstream genes through a feed-forward loop.EPXB5, encoding a beta-expansin family protein, is another direct target of TDF1, and it is highly expressed in the tapetum and pollen grains. The TDF1-AMS complex acts in concert to activate EXPB5 expression through a feed-forward loop. The identification of the regulatory pathway between TDF1 and AMS provides an interlocked feed-forward loop circuit that precisely regulates the transcriptional cascades that support anther development.

Background During anther development, the tapetum provides essential nutrients and materials for pollen development. In rice, multiple transcription factors and enzymes essential for tapetum development and pollen wall formation have been cloned from male-sterile lines. Results In this study, we obtained several lines in which the MYB transcription factor OsMS188 was knocked out through the CRISPR-Cas9 approach. The osms188 lines exhibited a male-sterile phenotype with aberrant development and degeneration of tapetal cells, absence of the sexine layer and defective anther cuticles. CYP703A3, CYP704B2, OsPKS1, OsPKS2, DPW and ABCG15 are sporopollenin synthesis and transport-related genes in rice. Plants with mutations in these genes are male sterile, with a defective sexine layer and anther cuticle. Further biochemical assays demonstrated that OsMS188 binds directly to the promoters of these genes to regulate their expression. UDT1, OsTDF1, TDR, bHLH142 and EAT1 are upstream regulators of rice tapetum development. Electrophoretic mobility shift assays (EMSAs) and activation assays revealed that TDR directly regulates OsMS188 expression. Additionally, protein interaction assays indicated that TDR interacts with OsMS188 to regulate downstream gene expression. Conclusion Overall, OsMS188 is a key regulator of tapetum development and pollen wall formation. The gene regulatory network established in this work may facilitate future investigations of fertility regulation in rice and in other crop species.