Nanobody-mediated control of gene expression and epigenetic memory

Nat Commun. 2021 Jan 22;12(1):537. doi: 10.1038/s41467-020-20757-1.

Abstract

Targeting chromatin regulators to specific genomic locations for gene control is emerging as a powerful method in basic research and synthetic biology. However, many chromatin regulators are large, making them difficult to deliver and combine in mammalian cells. Here, we develop a strategy for gene control using small nanobodies that bind and recruit endogenous chromatin regulators to a gene. We show that an antiGFP nanobody can be used to simultaneously visualize GFP-tagged chromatin regulators and control gene expression, and that nanobodies against HP1 and DNMT1 can silence a reporter gene. Moreover, combining nanobodies together or with other regulators, such as DNMT3A or KRAB, can enhance silencing speed and epigenetic memory. Finally, we use the slow silencing speed and high memory of antiDNMT1 to build a signal duration timer and recorder. These results set the basis for using nanobodies against chromatin regulators for controlling gene expression and epigenetic memory.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Animals
  • Chromatin / genetics
  • Chromatin / immunology*
  • Epigenesis, Genetic / immunology*
  • Gene Expression Regulation / immunology*
  • Gene Silencing / immunology
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / immunology
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Models, Genetic
  • Promoter Regions, Genetic / genetics
  • Promoter Regions, Genetic / immunology*
  • Signal Transduction / genetics
  • Signal Transduction / immunology
  • Single-Domain Antibodies / immunology*
  • Single-Domain Antibodies / metabolism

Substances

  • Chromatin
  • Single-Domain Antibodies
  • Green Fluorescent Proteins