Barcoded DNA nanostructures for the multiplexed profiling of subcellular protein distribution.
Nat Biomed Eng
; 3(9): 684-694, 2019 09.
Article
in En
| MEDLINE
| ID: mdl-31285580
ABSTRACT
Massively parallel DNA sequencing is established, yet high-throughput protein profiling remains challenging. Here, we report a barcoding approach that leverages the combinatorial sequence content and the configurational programmability of DNA nanostructures for high-throughput multiplexed profiling of the subcellular expression and distribution of proteins in whole cells. The barcodes are formed by in situ hybridization of tetrahedral DNA nanostructures and short DNA sequences conjugated with protein-targeting antibodies, and by nanostructure-assisted ligation (either enzymatic or chemical) of the nanostructures and exogenous DNA sequences bound to nanoparticles of different sizes (which cause these localization sequences to differentially distribute across subcellular compartments). Compared with linear DNA barcoding, the nanostructured barcodes enhance the signal by more than 100-fold. By implementing the barcoding approach on a microfluidic device for the analysis of rare patient samples, we show that molecular subtypes of breast cancer can be accurately classified and that subcellular spatial markers of disease aggressiveness can be identified.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
DNA
/
Gene Expression Profiling
/
Nanostructures
/
DNA Barcoding, Taxonomic
Limits:
Humans
Language:
En
Journal:
Nat Biomed Eng
Year:
2019
Document type:
Article
Affiliation country:
Singapore
Publication country:
ENGLAND
/
ESCOCIA
/
GB
/
GREAT BRITAIN
/
INGLATERRA
/
REINO UNIDO
/
SCOTLAND
/
UK
/
UNITED KINGDOM