High throughput PRIME editing screens identify functional DNA variants in the human genome

• Published in: bioRxiv : the preprint server for biology
• Main Authors: Ren, Xingjie, Yang, Han, Nierenberg, Jovia L, Sun, Yifan, Chen, Jiawen, Beaman, Cooper, Pham, Thu, Nobuhara, Mai, Takagi, Maya Asami, Narayan, Vivek, Li, Yun, Ziv, Elad, Shen, Yin
• Format: Journal Article
• Language: English
• Published: United States 12.07.2023

Despite tremendous progress in detecting DNA variants associated with human disease, interpreting their functional impact in a high-throughput and base-pair resolution manner remains challenging. Here, we develop a novel pooled prime editing screen method, PRIME, which can be applied to characterize thousands of coding and non-coding variants in a single experiment with high reproducibility. To showcase its applications, we first identified essential nucleotides for a 716 bp enhancer via PRIME-mediated saturation mutagenesis. Next, we applied PRIME to functionally characterize 1,304 non-coding variants associated with breast cancer and 3,699 variants from ClinVar. We discovered that 103 non-coding variants and 156 variants of uncertain significance are functional via affecting cell fitness. Collectively, we demonstrate PRIME capable of characterizing genetic variants at base-pair resolution and scale, advancing accurate genome annotation for disease risk prediction, diagnosis, and therapeutic target identification.