BiFC and FACS-based CRISPR screening revealed that QKI promotes PABPN1 LLPS in colorectal cancer cells

• DOI: 10.1093/procel/pwaf022
• Publication Date: 2025-03-07T11:46:22Z
• Authors: Mengxia Li; Zhijie Hu; Yingye Huang; Yuting Han; Cheng Liang; Yuchi Liu; Runze Wu; Xin Lu; Ke Deng; Susu Liu; Xin Ou; Yuwei Li; Chao Liu; Xuening Li; Jingting Liang; Yonggui Fu; Anlong Xu
• Keywords: Human genetics

Protein liquid-liquid phase separation (LLPS), a pivotal phenomenon intricately linked to cellular processes, is regulated by various other proteins. However, there still lack of high-throughput methods for screening protein regulators LLPS in target Here, we developed CRISPR/Cas9-based method identify integrating bimolecular fluorescence complementation (BiFC) and fluorescence-activated cell sorting (FACS). Using this newly method, screened the RNA-binding proteins that regulate PABPN1 identified tumor suppressor QKI as promoter separation. Furthermore, exhibits decreased expression levels diminished nuclear localization colorectal cancer cells, resulting reduced separation, which turn promotes alternative polyadenylation (APA), proliferation, migration cancer.