While technologies for measuring transcriptomes in single cells have matured, methods for measuring proteins and their post-translational modification (PTM) states in single cells are still being actively developed. Unlike nucleic acids, proteins cannot be amplified, making detection of minute quantities from single cells difficult. Here, we develop a strategy to detect targeted protein and its PTM isoforms in single cells. We barcode the proteins from single cells by tagging them with oligonucleotides, pool barcoded cells together, run bulk gel electrophoresis to separate protein and its PTM isoform and quantify their abundances by sequencing the oligonucleotides associated with each protein species. We used this strategy, iDentification and qUantification sEparaTion (DUET), to measure histone protein H2B and its monoubiquitination isoform, H2Bub, in single yeast cells. Our results revealed the heterogeneities of H2B ubiquitination levels in single cells from different cell-cycle stages, which is obscured in ensemble measurements.
|Publication status||Published - 2020 Dec 1|
Bibliographical noteFunding Information:
We thank R. Ismagilov for helpful suggestions and discussions. We thank I. Anto-schechkin from the Millard and Muriel Jacobs Genetics and Genomics Laboratory for assistance with the sequencing. We thank I. Strazhnik for help preparing the figures. This work was supported by funding from Paul G. Allen Frontiers Foundation Distinguished Investigator and Discovery Center.
© 2020, The Author(s).
All Science Journal Classification (ASJC) codes
- Medicine (miscellaneous)
- Biochemistry, Genetics and Molecular Biology(all)
- Agricultural and Biological Sciences(all)