A5097371909- CRISPR and Genetic Engineering
- RNA Interference and Gene Delivery
- Advanced biosensing and bioanalysis techniques
- RNA modifications and cancer
- RNA Research and Splicing
- Virus-based gene therapy research
University of Zurich
2022-2024
University Hospital of Zurich
2022
Abstract Arrayed CRISPR libraries extend the scope of gene-perturbation screens but require large numbers efficacious sgRNA-expressing vectors. Using a newly invented liquid-phase plasmid cloning methodology, we constructed genome-wide arrayed for human gene ablation (19,936 plasmids), activation, and epigenetic silencing (22,442 plasmids). At least 76% each preparation encoded an intact array 4 non-overlapping sgRNAs designed to tolerate most DNA polymorphisms. We achieved perturbation...
Abstract Arrayed CRISPR libraries extend the scope of gene-perturbation screens to non-selectable cell phenotypes. However, library generation requires assembling thousands vectors expressing single-guide RNAs (sgRNAs). Here, by leveraging massively parallel plasmid-cloning methodology, we show that arrayed can be constructed for genome-wide ablation (19,936 plasmids) human protein-coding genes and their activation epigenetic silencing (22,442 plasmids), with each plasmid encoding an array...
Abstract Heterogeneous Nuclear Ribonucleoprotein K (hnRNP K) is a limiting factor for prion propagation. However, little known about the function of hnRNP except that it essential to cell survival. Here, we performed synthetic-viability CRISPR ablation screen identify epistatic interactors HNRNPK . We found deletion Transcription Factor AP-2γ ( TFAP2C ) suppressed death K-depleted LN-229 and U-251 MG cells, whereas its overexpression hypersensitized cells loss. decreased cellular ATP,...