Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

TuNR: Orthogonal Control of Mean and Variability of Endogenous Genes in a Human Cell Line

View ORCID ProfileAlain R. Bonny, View ORCID ProfileJoão Pedro Fonseca, Jesslyn E. Park, View ORCID ProfileHana El-Samad
doi: https://doi.org/10.1101/2020.05.23.112979
Alain R. Bonny
1Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94158
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Alain R. Bonny
João Pedro Fonseca
1Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94158
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for João Pedro Fonseca
Jesslyn E. Park
1Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94158
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hana El-Samad
1Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94158
2Chan Zuckerberg Biohub, San Francisco, CA 94158
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Hana El-Samad
  • For correspondence: Hana.El-Samad@ucsf.edu
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Data/Code
  • Preview PDF
Loading

Abstract

Stochastic fluctuations at the transcriptional level contribute to isogenic cell-to-cell heterogeneity in mammalian cell populations. However, we still have no clear understanding of the repercussions of this heterogeneity, given the lack of tools to independently control mean expression and variability of a gene. Here, we engineered a synthetic circuit to independently modulate mean expression and heterogeneity of transgenes and endogenous human genes. The circuit, a Tunable Noise Rheostat (TuNR), consists of a transcriptional cascade of two inducible transcriptional activators, where the output mean and variance can be modulated by two orthogonal small molecule inputs. In this fashion, different combinations of the inputs can achieve the same mean but with different population variability. With TuNR, we achieve low basal expression, over 1000-fold expression of a transgene product, and up to 7-fold induction of the endogenous gene NGFR. Importantly, for the same mean expression level, we are able to establish varying degrees of heterogeneity in expression within an isogenic population, thereby decoupling gene expression noise from its mean. TuNR is therefore a modular tool that can be used in mammalian cells to enable direct interrogation of the implications of cell-to-cell variability.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • https://github.com/arb5134/Rheostat

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
Back to top
PreviousNext
Posted May 24, 2020.
Download PDF

Supplementary Material

Data/Code
Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
TuNR: Orthogonal Control of Mean and Variability of Endogenous Genes in a Human Cell Line
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
TuNR: Orthogonal Control of Mean and Variability of Endogenous Genes in a Human Cell Line
Alain R. Bonny, João Pedro Fonseca, Jesslyn E. Park, Hana El-Samad
bioRxiv 2020.05.23.112979; doi: https://doi.org/10.1101/2020.05.23.112979
Digg logo Reddit logo Twitter logo Facebook logo Google logo LinkedIn logo Mendeley logo
Citation Tools
TuNR: Orthogonal Control of Mean and Variability of Endogenous Genes in a Human Cell Line
Alain R. Bonny, João Pedro Fonseca, Jesslyn E. Park, Hana El-Samad
bioRxiv 2020.05.23.112979; doi: https://doi.org/10.1101/2020.05.23.112979

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Synthetic Biology
Subject Areas
All Articles
  • Animal Behavior and Cognition (3691)
  • Biochemistry (7800)
  • Bioengineering (5678)
  • Bioinformatics (21295)
  • Biophysics (10584)
  • Cancer Biology (8179)
  • Cell Biology (11948)
  • Clinical Trials (138)
  • Developmental Biology (6764)
  • Ecology (10401)
  • Epidemiology (2065)
  • Evolutionary Biology (13876)
  • Genetics (9709)
  • Genomics (13075)
  • Immunology (8151)
  • Microbiology (20022)
  • Molecular Biology (7859)
  • Neuroscience (43073)
  • Paleontology (321)
  • Pathology (1279)
  • Pharmacology and Toxicology (2261)
  • Physiology (3353)
  • Plant Biology (7232)
  • Scientific Communication and Education (1314)
  • Synthetic Biology (2008)
  • Systems Biology (5539)
  • Zoology (1128)