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Electrophoretic Analysis of Replication Through Structure-Prone DNA Repeats Within the SV40-Based Human Episome.
J Vis Exp
September 2024
Department of Biology, Tufts University.
Two-dimensional neutral/neutral gel-electrophoresis (2DGE) emerged as a benchmark technique to analyze DNA replication through natural impediments. This protocol describes how to analyze replication fork progression through structure-prone, expandable DNA repeats within the simian virus 40 (SV40)-based episome in human cells. In brief, upon plasmid transfection into human cells, replication intermediates are isolated by the modified Hirt protocol and treated with the DpnI restriction enzyme to remove non-replicated DNA.
View Article and Find Full Text PDFHistamine-releasing factor in severe asthma and rhinovirus-associated asthma exacerbation.
J Allergy Clin Immunol
September 2023
Laboratory of Allergic Diseases, La Jolla Institute for Immunology, La Jolla, Calif. Electronic address:
Background: Histamine-releasing factor (HRF) is implicated in allergic diseases. We previously showed its pathogenic role in murine models of asthma.
Objective: We aim to present data analysis from 3 separate human samples (sera samples from asthmatic patients, nasal washings from rhinovirus [RV]-infected individuals, and sera samples from patients with RV-induced asthma exacerbation) and 1 mouse sample to investigate correlates of HRF function in asthma and virus-induced asthma exacerbations.
Electrotaxis of alveolar epithelial cells in direct-current electric fields.
Chin J Traumatol
May 2023
Department of Respiratory Medicine, Daping Hospital, Third Military Medical University (Army Medical University), Chongqing, 400042, China; State Key Laboratory of Trauma, Burns, and Combined Injury, Department of Trauma Medical Center, Daping Hospital, Third Military Medical University (Army Medical University), Chongqing, 400042, China. Electronic address:
Purpose: This study aims to elucidate the electrotaxis response of alveolar epithelial cells (AECs) in direct-current electric fields (EFs), explore the impact of EFs on the cell fate of AECs, and lay the foundation for future exploitation of EFs for the treatment of acute lung injury.
Methods: AECs were extracted from rat lung tissues using magnetic-activated cell sorting. To elucidate the electrotaxis responses of AECs, different voltages of EFs (0, 50, 100, and 200 mV/mm) were applied to two types of AECs, respectively.
Comprehensive mutational analysis of the checkpoint signaling function of Rpa1/Ssb1 in fission yeast.
bioRxiv
March 2023
Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, Illinois, USA.
Unlabelled: Replication protein A (RPA) is a heterotrimeric complex and the major single-strand DNA (ssDNA) binding protein in eukaryotes. It plays important roles in DNA replication, repair, recombination, telomere maintenance, and checkpoint signaling. Because RPA is essential for cell survival, understanding its checkpoint signaling function in cells has been challenging.
View Article and Find Full Text PDFEfficient and reversible Cas13d-mediated knockdown with an all-in-one lentivirus-vector.
Front Bioeng Biotechnol
September 2022
Department of Biochemistry and Molecular Biology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Type VI CRISPR effector Cas13d from Ruminococcus flavefaciens XPD3002 (RfxCas13d) is an RNA-guided RNA endonuclease. RfxCas13d has been harnessed to knockdown gene expression with high specificity in various systems including mammalian cells. While inducible knockdown is advantageous over constitutive knockdown in many scenarios, current inducible systems of RfxCas13d express CRISPR RNA and Cas13d separately.
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