Identification of functional cis-regulatory elements by sequential enrichment from a randomized synthetic DNA library

BACKGROUND: The identification of endogenous cis-regulatory DNA elements (CREs) responsive to endogenous and environmental cues is important for studying gene regulation and for biotechnological applications but is labor and time intensive. Alternatively, by taking a synthetic biology approach small specific DNA binding sites tailored to the needs of the scientist can be generated and rapidly identified. RESULTS: Here we report a novel approach to identify stimulus-responsive synthetic CREs (SynCREs) from an unbiased random synthetic element (SynE) library. Functional SynCREs were isolated by screening the SynE libray for elements mediating transcriptional activity in plant protoplasts. Responsive elements were chromatin immunoprecipitated by targeting the active Ser-5 phosphorylated RNA polymerase II CTD (Pol II ChIP). Using sequential enrichment, deep sequencing and a bioinformatics pipeline, candidate responsive SynCREs were identified within a pool of constitutively active DNA elements and further validated. These included bonafide biotic/abiotic stress-responsive motifs along with novel SynCREs. We tested several SynCREs in Arabidopsis and confirmed their response to biotic stimuli. CONCLUSIONS: Successful isolation of synthetic stress-responsive elements from our screen illustrates the power of the described methodology. This approach can be applied to any transfectable eukaryotic system since it exploits a universal feature of the eukaryotic Pol II

Development of an Additively Manufactured Capacitive Humidity Sensor for the International Space Station

The ability to measure humidity on the International Space Station and other long-duration spaceflight missions is a crucial part of the onboard systems. For example, the Environmental Control and Life Support System (ECLSS) needs to know the amount of humidity in the air to make decisions about whether it should spend power to run the dehumidifier systems to attempt to reclaim that water. Other issues can arise if the humidity reaches too high of levels and condensation builds up on electrical components. With that in mind, it is vital that the spacecraft keeps spare sensors on board or has the ability to manufacture new sensors on demand. An additively manufactured sensor would be additionally beneficial because it would save space onboard that would normally be taken up by spares, save money from costly resupply missions, and allow the sensor to be constantly updated with the most effective design. This Technical Memorandum outlines a development process carried out to design, manufacture, and test an additively manufactured humidity sensor

Examining the Perspectives of Elementary Education Teachers Prepared Through Traditional and Dual License Programs

Preparing classroom teachers to work with students with diverse learning needs is a challenge that has been well documented by the literature. Earning a dual license in general and special education has been posited as one possible solution to this challenge. This paper reports on a qualitative study that examined the differences between dually licensed and traditionally prepared educators with regards to their self-efficacy and ideas about inclusion. Findings suggest that teachers who earn a dual license in general education and special education may have a stronger sense of self-efficacy as well as a stronger skill set for working with students with disabilities and other types of difference

Building Self-reliance: A Framework to Evaluate Smallholder Coffee Farmers’ Pursuit of Commercialization

This study examines the application of a self-reliance framework for practitioners and evaluators to better understand the capacities and intrinsic factors impacting smallholder coffee farmers’ commercialization behaviors. We surveyed 40 smallholder coffee producers in Peru using a quantitative instrument. Data were analyzed to determine if statistical relationships exist between farmers’ self-reliance (measured via knowledge and skills, attitudes, and aspirations) and their commercialization behaviors. Findings indicate the self-reliance framework effectively illustrates relationships between farmers’ aspirations, knowledge and skills and their commercialization behaviors, while future, additional studies are needed to better measure and understand the role of commercialization-related attitudes. Practitioners can leverage the study’s findings by using a self-reliance framework to infer farmers’ likeliness to pursue sustainable commercialization practices and align their trainings and design interventions based on evaluation findings. The conceptual self-reliance framework is the first of its kind applied for smallholder coffee commercialization. The findings demonstrate that self-reliance concepts employed recently in other contexts may potentially be used similarly by extension and development facilitators