US 12,281,301 B2
Sequencing-based proteomics
Feng Zhang, Cambridge, MA (US); Jonathan Leo Schmid-Burgk, Cambridge, MA (US); and Veit Hornung, Cambridge, MA (US)
Assigned to THE BROAD INSTITUTE, INC., Cambridge, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
Appl. No. 17/048,369
Filed by THE BROAD INSTITUTE, INC., Cambridge, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
PCT Filed Apr. 26, 2019, PCT No. PCT/US2019/029488
§ 371(c)(1), (2) Date Oct. 16, 2020,
PCT Pub. No. WO2019/210268, PCT Pub. Date Oct. 31, 2019.
Claims priority of provisional application 62/663,712, filed on Apr. 27, 2018.
Claims priority of provisional application 62/751,314, filed on Oct. 26, 2018.
Prior Publication US 2021/0147831 A1, May 20, 2021
Int. Cl. C40B 30/04 (2006.01); C12N 9/22 (2006.01); C12N 15/10 (2006.01); C12N 15/65 (2006.01)
CPC C12N 15/1065 (2013.01) [C12N 9/22 (2013.01); C12N 15/65 (2013.01); C12N 2310/20 (2017.05)] 17 Claims
OG exemplary drawing
 
1. A method of determining the distribution of protein levels in a population of cells, said method comprising:
a) sorting a library of cells into at least two groups based on expression of the detectable marker in each cell; and
b) nucleic acid sequencing of the cells in each group, wherein the tagged target genes in each group are determined;
wherein the library of cells comprises a plurality of cells, wherein each cell comprises a polynucleotide sequence encoding a protein tag integrated into the genome of the cell in frame with a protein coding gene selected from a set of target genes, wherein the library comprises more than one cell tagged at each target gene;
wherein the library of cells is treated with a perturbation prior to determining expression of proteins, wherein the perturbation comprises a small molecule, protein, RNAi, CRISPR system, TALE system, Zn finger system, meganuclease, pathogen, allergen, recombinant virus, temperature, salt, lipid, biomolecule, a pool of any perturbation thereof, or any combination thereof; and
wherein the protein tag is a detectable marker and the cell library is for use in detecting the distribution of protein levels in single cells comprising a plurality of cells.