Project Summary/Abstract: The past 20 years have seen a rapid expansion in the use of viral gene transfer vectors, with approved therapies and late stage clinical trials underway for the treatment of genetic disorders and multiple forms of cancer, as well as prevention of infectious diseases through vaccination. Major innovations in vector design and virus production have been accomplished for the three most widely used viral vector systems based on adenovirus, adeno- associated virus (AAV), and lentivirus. For laboratory investigators, cell and molecular biology methods to stably over-express and knockout a gene in cells and tissues have become indispensable in modern biomedical research. Lentivirus-mediated over-expression and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) knockout techniques are particularly powerful due to their efficiency and the capability of infecting dividing and non-dividing cells. Given the significant need and demand to use these viral gene transfer technologies and the lack of expert service providers in Rhode Island and the rest Southern New England region, we propose a Lentivirus Construct Core for the Stem Cells and Aging (SCA) COBRE Phase 3. Our long-term goal is to provide cutting-edge viral gene transfer technologies to the greater biomedical research community in Rhode Island and beyond. To accomplish this goal, we propose the following 4 Specific Aims: Specific Aim 1. To provide lentivirus technologies for easy access and efficient use. Specific Aim 2. To enhance the competitiveness of Rhode Island investigators to secure federal research funding. Specific Aim 3. To align our Core with translational research. Specific Aim 4. To become an independent self-sustainable service research facility. Innovations and impact: Recombinant viral vectors are powerful gene delivery tools for cells, animal models, and clinical applications. The lentiviral constructs from our Core will differ in their suitability for different applications, and will allow investigators to monitor cell functions, replace, correct, express or block expression of target genes, tag cells for fate determination, and change the physiological state of specific cell populations. The timely development of COVID-19 pseudovirus variants by our Core was a prime example of innovation. To genetically engineer oncolytic adenovirus encoding bispecific T cell engagers is cutting-edge, and the novel immunovirotherapies have the potential to make a profound impact in cancer treatments. The current exponential growth of clinical trials using AAV vectors suggests that we are only at the beginning of what is achievable for AAV as the leading platform for gene therapies. These innovations can potentially address diseases that have no other treatment options. In this vein, the Lentivirus Construct Core has already successfully made and will continue to make a positive impact as a catalyst on basic and translational research to improve human healt...