Biodistribution and Transcript Quantification Study in Mice

This study is a preclinical investigation aimed at characterizing the biodistribution and transgene expression of an AAV vector-delivered therapeutic in mouse tissues. The study utilized validated droplet digital PCR (ddPCR) and reverse transcription ddPCR (RT-ddPCR) methods to quantify vector copy numbers and vector-derived transcript copy numbers across various tissues. The objective was to support preclinical safety and efficacy assessments by determining tissue-specific distribution and transcriptional activity of the AAV vector. The methods involved optimizing assays on the Bio-Rad Droplet Digital PCR system, with quality controls prepared using naïve mouse tissues. The analysis included multiple tissues such as blood, brain, heart, and liver, confirming widespread biodistribution and active transcription of the AAV vector. The results indicated that the validated methods enabled sensitive and specific quantification of AAV vector DNA and transgene RNA, providing critical data for the preclinical evaluation of the therapeutic.