Nanobody-tethered transposition enables multifactorial chromatin profiling at single-cell resolution.
Nat Biotechnol
; 41(6): 806-812, 2023 06.
Article
em En
| MEDLINE
| ID: mdl-36536150
ABSTRACT
Chromatin states are functionally defined by a complex combination of histone modifications, transcription factor binding, DNA accessibility and other factors. Current methods for defining chromatin states cannot measure more than one aspect in a single experiment at single-cell resolution. Here we introduce nanobody-tethered transposition followed by sequencing (NTT-seq), an assay capable of measuring the genome-wide presence of up to three histone modifications and protein-DNA binding sites at single-cell resolution. NTT-seq uses recombinant Tn5 transposase fused to a set of secondary nanobodies (nb). Each nb-Tn5 fusion protein specifically binds to different immunoglobulin-G antibodies, enabling a mixture of primary antibodies binding different epitopes to be used in a single experiment. We apply bulk-cell and single-cell NTT-seq to generate high-resolution multimodal maps of chromatin states in cell culture and in human immune cells. We also extend NTT-seq to enable simultaneous profiling of cell surface protein expression and multimodal chromatin states to study cells of the immune system.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Cromatina
Limite:
Humans
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article