The sound of silence: transgene silencing in mammalian cell engineering

• Type of Material: Article
• Authors: Alan Cabera, Rice University; Hailey I. Edelstein, Northwestern University; Fokion Glykofrydis, University of Southern California; Kasey S. Love, Massachusetts Institute of Technology; Sebastian Palacios, Massachusetts Institute of Technology; Josh Tycko, Stanford University; Meng Zhang, University of Illinois Urbana-Champaign; Sarah Lensch, Stanford University; Cara E. Shields, Emory University; Mark Livingston, University of Utah; Ron Weiss, Massachusetts Institute of Technology; Huimin Zhao, University of Illinois Urbana-Champaign; Karmella Ann Haynes, Emory University; Leonardo Morsut, University of Southern California; Yvonne Y. Chen, University of California Los Angeles; Ahmad S. Khalil, Boston University; Wilson W. Wong, Boston University; James J. Collins, Massachusetts Institute of Technology; Susan J. Rosser, University of Edinburgh; Karen Polizzi, Imperial College London; Michael B. Elowitz, California Institute of Technology; Martin Fussenegger, ETH Zurich; Isaac B. Hilton, Rice University; Joshua N. Leonard, Northwestern University; Lacramioara Bintu, Stanford University; Kate E. Galloway, Massachusetts Institute of Technology; Tara L. Deans, University of Utah
• Language: English
• Date: 2022-12-21
• Publisher: Elsevier
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2022 The Authors. Published by Elsevier Inc.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): https://doi.org/10.1016/j.cels.2022.11.005
• Title of Journal or Parent Work: Cell Systems
• Volume: 13
• Issue: 12
• Start Page: 950
• End Page: 973
• Grant/Funding Information: Funding for this work was supported in part by the National Institutes of Health grants 1DP2CA250006-01 (T.L.D.), 1R01GM129011 (W.W.W.), R01EB029483 (W.W.W.), 1R01EB026510 (J.N.L), R21EB030772 (I.B.H.), R35GM143532 (I.B.H), R35GM143033 (K.E.G), R35GM138256 (L.M.), 4K00DK126120-03 (J.T.), R01EB029483 (A.S.K.), R35GM128947 (L.B.), R01EB030946 (R.W.), R01EB025256 (R.W.), 1UM1HG009402 (H.Z.), U54DK107965 (H.Z.), R21CA232244 (K.A.H.), 1RC2DK120535-01A1 (J.J.C.), and 1 U01 DK 127420- 01 (M.B.E.), the National Science Foundation grants CBET-2034495 (L.M.), CBET-2145528 (L.M.), 2141064 (K.S.L.), EF-1921677 (A.S.K) and EF-2021552 under subaward UWSC10142 (M.B.E.). Further support was also provided by the Biotechnology and Biological Sciences Research Council (BBSRC) BB/S006206/1 (K.P.) and BB/M018040/1 (S.J.R.), ElectroGene 785800 (M.F.), the SNF (M.F.), the Paul Allen Foundation (W.W.W.), AOFSR FA9550-22-1-0316 (K.E.G.), the Wellcome Sanger Institute LEAP 21-275 (L.M.), the Parker Institute for Cancer Immunotherapy (Y.Y.C.), the W.H. Coulter Department of Biomedical Engineering at Emory University (C.S.), and the DoD Vannevar Bush Faculty Fellowship N00014-20-1-2825 (A.S.K.). M.B.E. is a Howard Hughes Medical Institute Investigator.
• Keywords: transgene silencing; genome engineering; mammalian synthetic biology; synthetic gene circuit stability
• Research Categories: Biology, Genetics; Biology, Cell

Abstract

To elucidate principles operating in native biological systems and to develop novel biotechnologies, synthetic biology aims to build and integrate synthetic gene circuits within native transcriptional networks. The utility of synthetic gene circuits for cell engineering relies on the ability to control the expression of all constituent transgene components. Transgene silencing, defined as the loss of expression over time, persists as an obstacle for engineering primary cells and stem cells with transgenic cargos. In this review, we highlight the challenge that transgene silencing poses to the robust engineering of mammalian cells, outline potential molecular mechanisms of silencing, and present approaches for preventing transgene silencing. We conclude with a perspective identifying future research directions for improving the performance of synthetic gene circuits.