The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
Aug. 22, 2023

Filed:

Oct. 23, 2018
Applicants:

Massachusetts Institute of Technology, Cambridge, MA (US);

The General Hospital Corporation, Boston, MA (US);

Inventors:

Peter van Galen, Boston, MA (US);

Volker Hovestadt, Boston, MA (US);

Travis Hughes, Cambridge, MA (US);

Marc H. Wadsworth, II, Cambridge, MA (US);

Bradley Bernstein, Boston, MA (US);

Alexander K. Shalek, Cambridge, MA (US);

Todd M. Gierahn, Cambridge, MA (US);

J. Christopher Love, Cambridge, MA (US);

Ang A. Tu, Cambridge, MA (US);

Assignees:
Attorneys:
Primary Examiner:
Int. Cl.
CPC ...
C12P 19/34 (2006.01); C12N 15/10 (2006.01); C12Q 1/686 (2018.01); C12Q 1/6886 (2018.01); C12Q 1/6888 (2018.01);
U.S. Cl.
CPC ...
C12N 15/1065 (2013.01); C12Q 1/686 (2013.01); C12Q 1/6886 (2013.01); C12Q 1/6888 (2013.01); C12Q 2600/158 (2013.01);
Abstract

The present invention relates to methods of detecting region(s) of interest in a gene comprising a polyA tail. The region(s) of interest can include gene(s), region(s), mutation(s), deletion(s), insertion(s), indel(s), and/or translocation(s). The region(s) can be greater than or less than 1 kilobases from the polyA tail. Methods can include forming a library of single cell transcripts comprising the region(s) in close proximity to a cell barcode and a unique molecular identifier (UMI). Methods for distinguishing cells by genotype can include amplifying the transcripts using PCR methods and detecting the cell barcode and UMI using single cell sequencing methods. Transcripts can be enriched using tagged region-specific PCR primers. Cell barcodes can be brought into close proximity to the region(s) by circularizing the transcripts. Sequencing of the transcripts can include using primer binding sites added during PCR amplification and library indexes for multiplexed sequencing.


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