Efficient and Precise duplication and Inversion of Genomic DNA from Large to Chromosomal Scale

Genomic duplication is a fundamental force in molecular evolution and a major contributor to genetic and complex diseases. Chromosomal inversion, another prevalent form of structural variation, profoundly influences cellular fitness and genome stability. Here, we introduce two genome engineering platforms: amplification editing (AE), which enables programmable DNA duplication, and Prime Editing-based Inversion with Enhanced Performance (PIE), which achieves efficient induction of large-scale inversions in mammalian cells. AE supports the duplication of human genomic regions from as small as 20 base pairs to as large as 100 megabases (Mb), a scale comparable to entire human chromosomes. Using PIE, we reconfigured human chromosomes from metacentric to telocentric by inverting segments of 30 Mb and 100 Mb. Together, these technologies establish powerful approaches for engineering chromosomal structural variations, with broad applications in both medicine and biotechnology.

Learning Objectives:

1. Define the roles of genomic duplication and chromosomal inversion in evolution, genome stability, and disease.

2. Explain how amplification editing (AE) enables programmable DNA duplication across a wide range of scales.

3. Describe how Prime Editing-based Inversion with Enhanced Performance (PIE) reconfigures human chromosomes through large-scale inversions.