School of Science Division of Life Science 28 Genome Editing by CRISPR Supervisor: LIU Zhen / LIFS Student: LI Xingrui / BCB-IRE Course: UROP 2100, Fall UROP 3200, Summer This study investigates the use of CRISPR-Cas9 gene-editing technology to create cell models with targeted knockouts of genes related to cilia. Cilia are crucial for various cellular functions, and defects in motile cilia can lead to disorders such as Primary Ciliary Dyskinesia (PCD). By generating genetically modified HNCs or HBECs, we aim to explore the molecular mechanisms behind cilia-associated diseases. Through efficient CRISPR gene delivery and knockout validation, we observed significant disruptions in ciliary structure and function. High-speed video microscopy and immunostaining analysis further revealed the impact of specific gene knockouts on ciliary morphology and movement. This research provides valuable insights into the pathophysiology of cilia-related diseases and highlights CRISPR technology’s potential for studying complex genetic disorders. Genome Editing by CRISPR Supervisor: LIU Zhen / LIFS Student: TJANDRA Claresta Audrey / BIBU Course: UROP 1100, Spring Cilia are hair-like organelles essential for cellular signaling and sensory functions, with defects leading to ciliopathies that affect multiple organs. Primary ciliary dyskinesia (PCD), a recessive disorder caused by mutations in genes critical for ciliary components like dynein arms or radial spokes, disrupts motility and leads to chronic respiratory complications. This project uses CRISPR-Cas9 to knockout PCD-associated genes, utilizing custom-designed guide RNAs (gRNAs) for targeted disruption. Experimental workflows include gRNA design, annealing, ligation, and sequencing validation. By correlating genetic disruptions with altered ciliary beating dynamics, the study aims to establish definitive connections between specific mutations and PCD pathology, enhancing understanding of how structural defects drive clinical manifestations.
RkJQdWJsaXNoZXIy NDk5Njg=