The sound of silence: Transgene silencing in mammalian cell engineering

Creators: Cabrera, Alan; Edelstein, Hailey I.; Glykofrydis, Fokion; Love, Kasey S.; Palacios, Sebastian; Tycko, Josh; Zhang, Meng; Lensch, Sarah; Shields, Cara E.; Livingston, Mark; Weiss, Ron; Zhao, Huimin; Haynes, Karmella A.; Morsut, Leonardo; Chen, Yvonne Y.; Khalil, Ahmad S.; Wong, Wilson W.; Collins, James J.; Rosser, Susan J.; Polizzi, Karen; Elowitz, Michael B.¹; Fussenegger, Martin; Hilton, Isaac B.; Leonard, Joshua N.; Bintu, Lacramioara; Galloway, Kate E.; Deans, Tara L.

1. California Institute of Technology

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.

Additional Information

Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) 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 1U01 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.E.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. The authors listed with equal contribution are listed in the author list in alphabetical order.

Conflict of Interest

J.T. and L.B. acknowledge outside interest in Stylus Medicine.

Files

1-s2.0-S2405471222004641-main.pdf

Files (3.8 MB)

Name	Size	Download all
1-s2.0-S2405471222004641-main.pdf md5:6eb4cf74d0ded97c1d5f2d8e8b95eccf	3.8 MB	Preview Download

Additional details

	All versions	This version
Views	0	0
Downloads	0	0
Data volume	0 Bytes	0 Bytes