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:
Dec. 10, 2024

Filed:

Jun. 16, 2023
Applicant:

Insitro, Inc., South San Francisco, CA (US);

Inventors:

Cynthia Hao, Saratoga, CA (US);

Max R. Salick, San Francisco, CA (US);

Ci Chu, Palo Alto, CA (US);

Assignee:

Insitro, Inc., South San Francisco, CA (US);

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C12P 19/34 (2006.01); C12Q 1/6841 (2018.01); C12Q 1/6874 (2018.01); C12Q 1/6853 (2018.01);
U.S. Cl.
CPC ...
C12Q 1/6874 (2013.01); C12Q 1/6841 (2013.01); C12Q 1/6853 (2013.01);
Abstract

Disclosed herein are methods for performing in situ sequencing of RNA transcripts with non-uniform 5' ends. During reverse transcription (RT) of RNA transcripts, RT enzyme is induced to 'template-switch' to a separate oligonucleotide provided as the template for the upstream flanking region. This flanking region is grafted onto the beginning of the cDNA, enabling padlock probe detection, rolling circle amplification, and fluorescent in situ sequencing. Overall, the disclosed method for in situ sequencing can be applicable for analyzing exogenously introduced transcripts (e.g., identifying and determining impact of a perturbation including a CRISPR perturbation or shRNA/siRNA/ASO perturbation), analyzing naturally occurring transcripts (e.g., measuring gene expression, detecting splicing events), and analyzing modified, naturally occurring transcripts (e.g., detecting mutations or gene edits).


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