In seeking to support diversity, one challenge lies in adequately supporting and assessing science cognitions in a writing-intensive Biochemistry laboratory course when highly engaged Asian English language learners (Asian ELLs) struggle to communicate and make novice errors in English. Because they may understand advanced science concepts, but are not being adequately assessed for their deeper scientific understanding, we sought and examined interventions. We hypothesized that inquiry strategies, scaffolded learning through peer evaluation, and individualized tools that build writing communication skills would increase confidence. To assess scientific thinking, Linguistic Inquiry Word Count (LIWC) software measured underlying analytic and cognitive features of writing despite grammatical errors. To determine whether interventions improved student experience or learning outcomes, we investigated a cross-sectional sample of cases within experimental groups (n = 19) using a mixed-methods approach. Overall trends of paired t-tests from Asian ELLs’ pre/post surveys showed gains in six measures of writing confidence, with some statistically significant gains in confidence in writing skill (p=0.025) and in theory (p≤0.05). LIWC scores for Asian ELL and native-English-speaking students were comparable except for increased cognitive scores for Asian ELLs and detectable individual differences. An increase in Asian ELLs’ cognitive scores in spring/summer over fall was observed (p = 0.04), likely as a result of greater cognitive processes with language use, inquiry-related interventions, and peer evaluation. Individual cases further elucidated challenges faced by Asian ELL students. LIWC scores of student writing may be useful in determining underlying understanding. Interventions designed to provide support and strengthen the writing of Asian ELL students may also improve their confidence in writing, even if improvement is gradual.

HUH‐endonucleases (histidine‐hydrophobic‐histidine) create nicks in hairpin loops at the origin of replication during viral rolling circle replication and bacterial plasmid conjugation. Intriguingly, these metal‐dependent enzymes can form rapid, covalent, and specific linkages with single‐stranded DNA (ssDNA) under physiologic conditions. In recent years, we have developed these nucleases as a multiplexable, protein‐DNA bioconjugation (HUH‐tags) technology for live cell imaging, force spectroscopy, genome editing, cell targeting, etc. Though viral HUH‐tags bear valuable biophysical properties for bioconjugation, it would seem multiplexing the viral HUH‐tags we explored would be difficult because they target a highly conserved nonamer sequence (5′‐TA(A/G)TATT*AC‐3′). Interestingly, we noticed varying degrees of promiscuity in sequence recognition between these HUH‐tags. Therefore, we hypothesized that we could exploit or engineer sequence orthogonality using variants of the target sequence. To this end, we generated an unbiased ssDNA library containing all permutations of the target sequence 5′ of the cleavage site (16,384 sequences) and developed a next‐generation sequencing approach that assesses the degree of HUH‐tag cleavage of all sequences contained in the library. Strikingly, we found robust sequence orthogonality and can now multiplex several of these viral HUH‐tags. Our ultimate aim is to engineer the target sequence recognition to create a vast toolkit of multiplexable, viral HUH‐tags. However, relatively few structural and mechanistic insights into the ssDNA recognition have limited our ability to rationally engineer HUH‐tags for this purpose. Addressing this bottleneck, we solved the first ever structure of a HUH‐tag derived from a viral replication initiator protein from wheat dwarf virus bound to its ssDNA target generating an atomic resolution map of protein‐DNA contacts. Together, our structural insights and sequence cleavage screening method may allow us to rationally develop and engineer a powerful toolkit of fusion tags for multiplexable protein‐DNA bioconjugation. Support or Funding Information NIH: R35GM119483, University of Minnesota

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Science concept recall and engagement take place through individual pre‐lab preparation, in‐lab partnerships, hands‐on training, and social learning communities within the Social Ecological Model and Cognitive Theory frameworks. STEM majors are introduced to key concepts and skills at several stages in pre‐requisite courses required for later enrollment in our Biochemistry Laboratory course. Because this curriculum assumes high recall, we measured student understanding and recall in a series of pre‐skill tests at the lab course onset: Polymerase Chain Reaction (PCR) ability to create potential primers for a strand of DNA; math dilution skills and ability to calculate molarity for a buffer solution; and other concepts assumed to be easily recalled for application to lab course exercises. Per common practice, we employed pre‐lab exercises to help refresh students' memories and ensure lab preparation. A surprising gap in understanding of PCR was detected, so pre‐lab interventions were investigated along with near‐peer mentoring in a co‐mentoring community group to promote learning inclusion.Spanning iterative semesters (Fall 2016–Fall 2018) of an upper‐level Biochemistry lab course, we used pre‐post assessments of de‐identified student data per IRB (sample size n=56–90 per semester) to investigate 1) recall, 2) iterative versions of pre‐lab prompts for enhanced skill and confidence, and 3) different student outcomes per intervention. Skill assessments were qualitatively coded and hand‐scored for correctness, analyzed, and matched to paired pre‐post student self‐report in a 5‐point Likert scale of confidence in theory and confidence in skill: Doing PCR, Designing PCR Primers, Translation of DNA, Dilution Calculations, Molarity Calculations, etc.Student recall on the first lab day varied widely in pre‐assessments. Confidence in the theory and skill trended with higher incoming self‐reported confidence than actual ability to design a primer, and higher confidence with theory than with skill. Results varied among no pre‐lab, paper‐based, and online links with videos suggesting overall increased success in skill and confidence measured in post‐course assessments, yet post‐course student outcomes were still suboptimal. An additional post‐course intervention recruiting students for a co‐mentoring community model with a more course‐based undergraduate research (CUR) approach provided deeper learning out of class, social support, and additional outcomes.Use of pre‐labs was valuable to promote recall; however, pre‐lab effectiveness required deeper prompts. Simply reading and doing PCR in lab was not sufficient, nor was repeated lecture, discussion, or quizzing. The co‐mentoring community model extended the course learning to more applied learning and broader impacts building STEM learning and inclusion.Support or Funding InformationUniversity of Minnesota Grant‐in‐AidThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.