Targeted and inducible regulation of mammalian gene expression is a broadly important capability. We engineered drug-inducible catalytically inactive Cpf1 nuclease fused to transcriptional activation domains to tune the expression of endogenous genes in human cells. Leveraging the multiplex capability of the Cpf1 platform, we demonstrate both synergistic and combinatorial gene expression in human cells. Our work should enable the development of multiplex gene perturbation library screens for understanding complex cellular phenotypes.
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http://dx.doi.org/10.1038/nmeth.4483 | DOI Listing |
J Inflamm Res
January 2025
Department of Orthopaedic Surgery, Third Hospital of Hebei Medical University, Shijiazhuang, Hebei, People's Republic of China.
Purpose: Necrotizing fasciitis (NF) is a scarce but potentially life-threatening infection. However, no research has reported the cellular heterogeneity in patients with NF. We aim to investigate the change of cells from deep fascia in response to NF by single-cell RNA-seq.
View Article and Find Full Text PDFMultiplexed assays of variant effect (MAVEs) perform simultaneous characterization of many variants. Prime editing has been recently adopted for introducing many variants in their native genomic contexts. However, robust protocols and standards are limited, preventing widespread uptake.
View Article and Find Full Text PDFHeliyon
January 2025
Department of Medicine, Sylhet Agricultural University, Sylhet, 3100, Bangladesh.
spp. present in the food chain have been of much interest during the last few decades due to their implication in the development of antimicrobial resistance. We determined the prevalence of spp.
View Article and Find Full Text PDFJ Vet Diagn Invest
January 2025
Asia-Pacific Centre for Animal Health, Melbourne Veterinary School, Faculty of Science, University of Melbourne, Werribee, Victoria, Australia.
The complex contains important opportunistic pathogens of humans and vertebrate animals, as well as insects and other invertebrates. To date, the methods used for the identification of species within the genus , including PCR assays, have poor discriminatory power and may require further molecular typing or genomic sequence analysis to determine clinical relevance. We developed a duplex TaqMan probe-based quantitative real-time PCR (qPCR) assay targeting the gene, which is involved in chitin degradation and transport, and the gene, which is involved in urease production.
View Article and Find Full Text PDFPlant Biotechnol J
January 2025
Wageningen University and Research, Plant Breeding, Wageningen, The Netherlands.
Mutants with simultaneous germline mutations were obtained in all three F5H genes and all three FAD2 genes (one to eleven mutated alleles) in order to improve the feed value of the seed meal and the fatty acid composition of the seed oil. In mutants with multiple mutated F5H alleles, sinapine in seed meal was reduced by up to 100%, accompanied by a sharp reduction in the S-monolignol content of lignin without causing lodging or stem break. A lower S-lignin monomer content in stems can contribute to improved stem degradability allowing new uses of stems.
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