3 articles published in JoVE
Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9 Lorenzo Brunetti*1,2,3, Michael C. Gundry*1,2,4, Ayumi Kitano4, Daisuke Nakada1,2,4, Margaret A. Goodell1,2,4,5 1Stem Cells & Regenerative Medicine Center, Baylor College of Medicine, 2Center for Cell and Gene Therapy, Baylor College of Medicine, 3Centro di Ricerca Emato-Oncologica (CREO), University of Perugia, 4Department of Molecular & Human Genetics, Baylor College of Medicine, 5 A protocol for fast CRISPR/Cas9-mediated gene disruption in mouse and human primary hematopoietic cells is described in this article. Cas9-sgRNA ribonucleoproteins are introduced via electroporation with sgRNAs generated through in vitro transcription and commercial Cas9. High editing efficiencies are achieved with limited time and financial cost.
Preparation of Extracellular Matrix Protein Fibers for Brillouin Spectroscopy Ryan S. Edginton1, Sara Mattana2, Silvia Caponi2,3, Daniele Fioretto2, Ellen Green1, C. Peter Winlove1, Francesca Palombo1 1School of Physics and Astronomy, University of Exeter, 2Department of Physics and Geology, University of Perugia, 3Istituto Officina dei Materiali del CNR, Unità di Perugia We present a protocol for the application of Brillouin light scattering spectroscopy to elastin and trypsin-purified type I collagen fibers of the extracellular matrix to extract their full elastic properties.
A Comparative Analysis of Recombinant Protein Expression in Different Biofactories: Bacteria, Insect Cells and Plant Systems Elisa Gecchele*1, Matilde Merlin*1, Annalisa Brozzetti2, Alberto Falorni2, Mario Pezzotti1, Linda Avesani1 1Department of Biotechnology, University of Verona, Verona, Italy, 2Department of Internal Medicine, University of Perugia, Perugia, Italy In this study the expression of a target human recombinant protein in different production platforms was compared. We focused on traditional fermenter-based cultures and on plants, describing the set-up of each system and highlighting, on the basis of the reported results, the inherent limits and advantages for each platform.