Targeted enrichment and high‐resolution digital profiling of mitochondrial DNA deletions in human brain

摘要: Due largely to the inability accurately quantify and characterize de novo deletion events, mechanisms underpinning pathogenic expansion of mtDNA deletions in aging neuromuscular disorders remain poorly understood. Here, we outline validate a new tool termed ‘Digital Deletion Detection’ (3D) that allows for high-resolution analysis rare occurring at frequencies as low 1 × 10−8. 3D is three-step process includes targeted enrichment deletion-bearing molecules, single-molecule partitioning genomes into thousands droplets direct quantification via droplet digital PCR, breakpoint characterization using massively parallel sequencing. Using 3D, interrogated over 8 billion mitochondrial analyze age-related dynamics human brain tissue. We demonstrate total load increases with age, while number diversity unique constant. Our data provide support hypothesis pre-existing mutations primary factor contributing accumulation deletions.