PAM-flexible genome editing with an engineered chimeric Cas9

CRISPR enzymes require a defined protospacer adjacent motif (PAM) flanking a guide RNA-programmed target site, limiting their sequence accessibility for robust genome editing applications. In this study, we recombine the PAM-interacting domain of SpRY, a broad-targeting Cas9 possessing an NRN > NYN (R = A or G, Y = C or T) PAM preference, with the N-terminus of Sc + +, a Cas9 with simultaneously broad, efficient, and accurate NNG editing capabilities, to generate a chimeric enzyme with highly flexible PAM preference: SpRYc. We demonstrate that SpRYc leverages properties of both enzymes to specifically edit diverse PAMs and disease-related loci for potential therapeutic applications. In total, the approaches to generate SpRYc, coupled with its robust flexibility, highlight the power of integrative protein design for Cas9 engineering and motivate downstream editing applications that require precise genomic positioning

A case-based approach for teaching professionalism to residents with online discussions

Introduction: Programs must demonstrate that their residents are taught and assessed in professionalism. Most programs struggle with finding viable ways to teach and assess this critical competency. UTHSCSA Family and Community Medicine Residency developed an innovative option for interactive learning and assessment of residents in this competency which would be transferrable to other programs and specialties. Methods: The innovative approach uses an asynchronous online format on Blackboard. Threaded discussions on Blackboard require thoughtful reflective writing after case assessment and critical evaluation of other resident posts. Participation, content and progress of all resident postings are monitored by administrative staff and faculty. Faculty can further engage the residents at any point to deepen the discussion and learning. Results: 100% of all senior residents attained the required learning objectives. All were actively engaged in the assignments. Six cases have been developed using a Learning Matrix to demonstrate evaluation areas from the specialty specific competencies. Written feedback from residents verified the validity of case content in context of their current clinical practice. Postings by residents have provided value and insight for the faculty to access the professional development of our Family Medicine residents. The Clinical Competency Committee evaluates all third year residents using this information specific to the professionalism milestones. By using an asynchronous online approach to case discussion, all residents are involved with all aspects of this curriculum. Conclusions: More specific measurable learning outcomes are possible using this approach. Resident participation and engagement is easier to track and monitor than a lecture-based format and easier to capture valuable data than relying on evaluation feedback. Our Annual Review process will identify areas for improvement in the existing cases and help create supplemental cases based on a needs assessment by the faculty

A Cas9 with PAM recognition for adenine dinucleotides

CRISPR-associated (Cas) DNA-endonucleases are remarkably effective tools for genome engineering, but have limited target ranges due to their protospacer adjacent motif (PAM) requirements. We demonstrate a critical expansion of the targetable sequence space for a type II-A CRISPR-associated enzyme through identification of the natural 5′-NAAN-3′ PAM preference of Streptococcus macacae Cas9 (SmacCas9). To achieve efficient editing activity, we graft the PAM-interacting domain of SmacCas9 to its well-established ortholog from Streptococcus pyogenes (SpyCas9), and further engineer an increased efficiency variant (iSpyMac) for robust genome editing activity. We establish that our hybrids can target all adenine dinucleotide PAM sequences and possess robust and accurate editing capabilities in human cells

Implementation of interprofessional education (IPE) in 16 U.S. medical schools: Common practices, barriers and facilitators.

BackgroundEnhanced patient outcomes and accreditation criteria have led schools to integrate interprofessional education (IPE). While several studies describe IPE curricula at individual institutions, few examine practices across multiple institutions.PurposeTo examine the IPE integration at different institutions and determine gaps where there is potential for improvement.MethodIn this mixed methods study, we obtained survey results from 16 U.S. medical schools, 14 of which reported IPE activities.ResultsThe most common collaboration was between medical and nursing schools (93%). The prevalent format was shared curriculum, often including integrated modules (57%). Small group activities represented the majority (64%) of event settings, and simulation-based learning, games and role-play (71%) were the most utilized learning methods. Thirteen schools (81.3%) reported teaching IPE competencies, but significant variation existed. Gaps and barriers in the study include limitations of using a convenience sample, limited qualitative analysis, and survey by self-report.ConclusionsMost IPE activities focused on the physician role. Implementation challenges included scheduling, logistics and financial support. A need for effective faculty development as well as measures to examine the link between IPE learning outcomes and patient outcomes were identified