Version 1
: Received: 1 December 2023 / Approved: 4 December 2023 / Online: 4 December 2023 (10:18:09 CET)
How to cite:
Chua, R.; Wang, L.; Singaraja, R.; Ghosh, S. Functional and Multi-Omics Analysis of an Optimized CRISPR-Mediated FURIN Depletion in Cultured Pro-Monocytic Cells. Preprints2023, 2023120178. https://doi.org/10.20944/preprints202312.0178.v1
Chua, R.; Wang, L.; Singaraja, R.; Ghosh, S. Functional and Multi-Omics Analysis of an Optimized CRISPR-Mediated FURIN Depletion in Cultured Pro-Monocytic Cells. Preprints 2023, 2023120178. https://doi.org/10.20944/preprints202312.0178.v1
Chua, R.; Wang, L.; Singaraja, R.; Ghosh, S. Functional and Multi-Omics Analysis of an Optimized CRISPR-Mediated FURIN Depletion in Cultured Pro-Monocytic Cells. Preprints2023, 2023120178. https://doi.org/10.20944/preprints202312.0178.v1
APA Style
Chua, R., Wang, L., Singaraja, R., & Ghosh, S. (2023). Functional and Multi-Omics Analysis of an Optimized CRISPR-Mediated FURIN Depletion in Cultured Pro-Monocytic Cells. Preprints. https://doi.org/10.20944/preprints202312.0178.v1
Chicago/Turabian Style
Chua, R., Roshni Singaraja and Sujoy Ghosh. 2023 "Functional and Multi-Omics Analysis of an Optimized CRISPR-Mediated FURIN Depletion in Cultured Pro-Monocytic Cells" Preprints. https://doi.org/10.20944/preprints202312.0178.v1
Abstract
The pro-protein convertase FURIN (PCSK3) is implicated in a wide range of normal and pathological biological processes such as infectious disease, cancer and cardiovascular disease. Previously, we performed a systemic inhibition of FURIN in a mouse model of atherosclerosis and demonstrated significant plaque reduction and alterations in macrophage function. To understand the cellular mechanisms affected by FURIN inhibition in myeloid cells, we optimized a CRISPR-mediated gene deletion protocol for successfully deriving hemizygous (HZ) and nullizygous (NZ) FURIN knockout clones in U937 monocytic cells using lipotransfection based procedures and a dual guide RNA delivery strategy. We observed differences in monocyte and macrophage functions involving phagocytosis, lipid accumulation, cell migration, inflammatory gene expression, cytokine release patterns, secreted proteomics (cytokines) and whole-genome transcriptomics between wild-type, HZ and NZ FURIN clones. These studies provide a mechanistic basis on the possible roles of myeloid cell FURIN in cardiovascular disorders.
Biology and Life Sciences, Cell and Developmental Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
