Rapid Generation of MicroRNA Sponges for MicroRNA Inhibition

MicroRNA (miRNA) sponges are transcripts with repeated miRNA antisense sequences that can sequester miRNAs from endogenous targets. MiRNA sponges are valuable tools for miRNA loss-of-function studies both in vitro and in vivo. We developed a fast and flexible method to generate miRNA sponges and tested their efficiency in various assays. Using a single directional ligation reaction we generated sponges with 10 or more miRNA binding sites. Luciferase and AGO2-immuno precipitation (IP) assays confirmed effective binding of the miRNAs to the sponges. Using a GFP competition assay we showed that miR-19 sponges with central mismatches in the miRNA binding sites are efficient miRNA inhibitors while sponges with perfect antisense binding sites are not. Quantification of miRNA sponge levels suggests that this is at least in part due to degradation of the perfect antisense sponge transcripts. Finally, we provide evidence that combined inhibition of miRNAs of the miR-17∼92 cluster results in a more effective growth inhibition as compared to inhibition of individual miRNAs. In conclusion, we describe and validate a method to rapidly generate miRNA sponges for miRNA loss-of-function studies

Using Feminist Theory and Social Justice Pedagogy to Educate a New Generation of Precautionary Principle Chemists

Students in the science classroom learn that science makes right, which devalues (whether inadvertently or very purposefully) other non-scientific areas of study and largely works against ideas of collectivism and community especially as science “rises to the top” in collaborative partnerships and interdisciplinary research and service. Applying feminist theory and social justice pedagogy within the chemistry classroom can help focus the student’s learning experiences away from hierarchical outcomes that perpetuate discriminatory practices toward change agency and the pursuit of equity, especially in regard to the design of safer chemicals. Most would agree that it is not ethically acceptable to continue to design and produce chemicals that cause adverse human health effects and environmental pollution; however, changes in the chemistry classroom to redirect this outcome has been slow. Emphasis on designing safer chemicals driven by the precautionary principle can advance the role of students and teachers as change agents in their communities.. When considering feminist theory and social justice pedagogy in the chemistry classroom, classrooms become inclusive and learning outcomes shift toward focusing on how to reduce health disparities due to chemical exposures and how to interact with systems of domination and decision making in the workplace and beyond. New chemistry curriculum is needed in these areas to help prepare a new generation of “precautionary principle chemists” who will advocate for all of our communities

Transitioning to Safer Chemicals in Academic Research Laboratories: Lessons Learned at the University of Washington

Chemicals are an integral component of laboratory activities in academia but minimizing hazards and environmental impacts of chemicals is challenging. This paper describes how laboratories in University of Washington’s Department of Environmental and Occupational Health Sciences (UW DEOHS) partnered with the UW Green Laboratory Program to explore approaches for assisting laboratories to adopt green chemistry principles and select safer chemicals. Chemical inventories, purchasing records, and hazardous waste data were used to quantitate chemical use in DEOHS. Characterizing chemical use based on the data sources provided the project team with a summary of the high volume chemicals used by departmental laboratories. As a way to target chemicals that are highly hazardous but not used in large masses/volumes, laboratory managers were asked about highly hazardous or toxic chemicals they used. Two chemicals were selected for alternatives assessments and developed into case studies that represent different barriers that laboratories face in their efforts to transition to greener and safer chemicals. This project provided a unique opportunity to survey chemicals used by a set of laboratories with diverse research topics and to assess the practicality of transitioning to safer and greener chemicals in laboratory research using case studies