Abstract 485: Using the Celigo™ Image Cytometer for effectively screening rAAV gene delivery methods

Abstract To develop effective rAAV gene delivery vectors, scientists need to consider multiple elements, such as the promoter, insertion location, genetic target, and target gene to name a few. Green fluorescent protein (GFP), or other detection markers are often used to measure vector transduction and gene expression efficiency. Transduced GFP samples can be analyzed using fluorescent microscopy and flow cytometry, however, both methods can be quite time consuming, labor intensive, and lack high-throughput capabilities to kinetically monitor gene expression. In this work, we developed and employed the Celigo™ Image Cytometer to screen different gene delivery vectors by comparing GFP expression level in SupT1 cells at 24-, 48-, 72-, and 96-hours following transduction. Using the Celigo™ we were able to monitor transient and integrated GFP expression amongst multiple gene delivery vectors. For transient expression monitoring, SB-P-AA-101-07 displayed stronger expression levels at 24- and 48-hours. For integrated expression monitoring, both plasmid and rAAV with integration element exhibited over ∼90%+ GFP transduction efficiency by 96-hours. In summary, the Celigo™ may prove useful for researchers developing and screening gene delivery vectors in a high throughput manner. Note: SB-P-AA-101-07 (from Revvity Health Sciences, Inc.) is for research use only. Not for use in diagnostic procedures. Citation Format: Mackenzie Pierce, Yongyang Huang, Bo Lin. Using the Celigo™ Image Cytometer for effectively screening rAAV gene delivery methods [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 485.