University of San Diego 3 articles published in JoVE Biology Reconstituting and Characterizing Actin-Microtubule Composites with Tunable Motor-Driven Dynamics and Mechanics Mehrzad Sasanpour1, Daisy H. Achiriloaie1,2, Gloria Lee1, Gregor Leech1, Maya Hendija1, K. Alice Lindsay3, Jennifer L. Ross3, Ryan J. McGorty1, Rae M. Robertson-Anderson1 1Department of Physics and Biophysics, University of San Diego, 2W. M. Keck Science Department, Scripps College, Pitzer College, and Claremont McKenna College, 3Department of Physics, Syracuse University This paper presents protocols for engineering and characterizing tunable three-dimensional composite networks of co-entangled actin filaments and microtubules. Composites undergo active restructuring and ballistic motion, driven by myosin II and kinesin motors, and are tuned by the relative concentrations of actin, microtubules, motor proteins, and passive crosslinkers. Bioengineering Quantifying Cytoskeleton Dynamics Using Differential Dynamic Microscopy Hannah N. Verwei1, Gloria Lee2, Gregor Leech2, Irene Istúriz Petitjean3, Gijsje H. Koenderink3, Rae M. Robertson-Anderson2, Ryan James McGorty2 1Cell Biology, Neurobiology and Biophysics, Department of Biology, Faculty of Science, Utrecht University, 2Department of Physics and Biophysics, University of San Diego, 3Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology Differential dynamic microscopy (DDM) combines features of dynamic light scattering and microscopy. Here, the process of using DDM to characterize reconstituted cytoskeleton networks by quantifying the subdiffusive and caged dynamics of particles in vimentin networks and the ballistic motion of active myosin-driven actin-microtubule composites is presented. Bioengineering Gel-seq: A Method for Simultaneous Sequencing Library Preparation of DNA and RNA Using Hydrogel Matrices Gordon D. Hoople*1, Andrew Richards*2, Yan Wu2, Albert P. Pisano3, Kun Zhang2 1Department of Engineering, University of San Diego, 2Department of Bioengineering, University of California, San Diego, 3Department of Mechanical and Aerospace Engineering, University of California, San Diego Gel-seq enables researchers to simultaneously prepare libraries for both DNA- and RNA-seq at negligible added cost starting from 100 - 1000 cells using a simple hydrogel device. This paper presents a detailed approach for the fabrication of the device as well as the biological protocol to generate paired libraries.
