A genome-scale resource for in vivo tag-based protein function exploration in C. elegans.

Understanding the in vivo dynamics of protein localization and their physical interactions is important for many problems in biology. To enable systematic protein function interrogation in a multicellular context, we built a genome-scale transgenic platform for in vivo expression of fluorescent- and affinity-tagged proteins in Caenorhabditis elegans under endogenous cis regulatory control. The platform combines computer-assisted transgene design, massively parallel DNA engineering, and next-generation sequencing to generate a resource of 14,637 genomic DNA transgenes, which covers 73% of the proteome. The multipurpose tag used allows any protein of interest to be localized in vivo or affinity purified using standard tag-based assays. We illustrate the utility of the resource by systematic chromatin immunopurification and automated 4D imaging, which produced detailed DNA binding and cell/tissue distribution maps for key transcription factor proteins.

Evaluation of Ongoing Crisis Intervention Team (CIT) Training for Law Enforcement Using the ECHO Model.

Crisis Intervention Team (CIT) training aims to improve law enforcement officers' (LEOs) ability to safely intervene in calls for service involving individuals with mental illness, as well as to increase LEOs' ability to link these individuals to mental health services and divert them from the criminal justice system. However, most CIT training is delivered as a stand-alone class, and continuing education in CIT principles and best practices is limited. To address this problem, the Albuquerque Police Department, in partnership with the Department of Psychiatry and Behavioral Sciences at the University of New Mexico, developed CIT ECHO to provide continuing education in CIT best practices. The authors evaluated 113 weekly CIT ECHO sessions targeting LEOs in New Mexico, offered between 2017 and 2020. LEOs electronically completed a post-session survey after each didactic; additionally, a targeted follow-up survey was distributed to LEOs participating in at least three sessions. Surveys measured impact of CIT ECHO on knowledge, self-efficacy, and attitudes towards individuals with mental illness involved in the criminal justice system. After participating in CIT ECHO, LEOs reported increases in knowledge of didactic content and that they felt comfortable applying didactic content on the job. LEOs also evidenced positive attitudinal shifts towards individuals with mental illness and criminal justice involvement. Continuing education in CIT best practices appears to increase LEOs' knowledge base and comfort in working with individuals with mental illness and criminal justice involvement, as well as results in positive attitudinal shifts towards this population.

Beyond crisis intervention team (CIT) classroom training: Videoconference continuing education for law enforcement.

Continuing education in Crisis Intervention Team (CIT) principles and best practices are limited. In 2015, the Albuquerque Police Department became the first law enforcement agency in the country to provide extended learning for CIT and case debriefings related to behavioral health through videoconferencing technology. The project, known as CIT ECHO, connects law enforcement agencies across New Mexico and the country to an online classroom where CIT experts and psychiatrists review behavioral health topics, and debrief complex cases with officers. An overview of CIT ECHO is provided including key elements, implementation challenges, and how we are evaluating its reach and effectiveness.

Rab5-family guanine nucleotide exchange factors bind retromer and promote its recruitment to endosomes.

The retromer complex facilitates the sorting of integral membrane proteins from the endosome to the late Golgi. In mammalian cells, the efficient recruitment of retromer to endosomes requires the lipid phosphatidylinositol 3-phosphate (PI3P) as well as Rab5 and Rab7 GTPases. However, in yeast, the role of Rabs in recruiting retromer to endosomes is less clear. We identified novel physical interactions between retromer and the Saccharomyces cerevisiae VPS9-domain Rab5-family guanine nucleotide exchange factors (GEFs) Muk1 and Vps9. Furthermore, we identified a new yeast VPS9 domain-containing protein, VARP-like 1 (Vrl1), which is related to the human VARP protein. All three VPS9 domain-containing proteins show localization to endosomes, and the presence of any one of them is necessary for the endosomal recruitment of retromer. We find that expression of an active VPS9-domain protein is required for correct localization of the phosphatidylinositol 3-kinase Vps34 and the production of endosomal PI3P. These results suggest that VPS9 GEFs promote retromer recruitment by establishing PI3P-enriched domains at the endosomal membrane. The interaction of retromer with distinct VPS9 GEFs could thus link GEF-dependent regulatory inputs to the temporal or spatial coordination of retromer assembly or function.