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US 2025/0084484 A1Published application
Methods and compositions for transcriptome analysis
AssigneeJumpcode Genomics, Inc.
InventorKeith BROWN
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Abstract
Provided herein are methods and compositions for analysis of a transcriptome, for example detection of a RNA isoform, of a cell, tissue, organ, or subject.
Claims20
A method of detecting an RNA isoform comprising:
- (a) contacting a plurality of RNA molecules to a oligo(dT) primer and a reverse transcriptase to produce a plurality of cDNA molecules;
- (b) attaching adaptors to a 5′ end and a 3′ end of each of said plurality of cDNA molecules;
- (c) amplifying said plurality of cDNA molecules with a first primer and a second primer that bind to said adaptors to produce a plurality of amplification products, wherein said first primer or said second primer comprises a detectable label;
- (d) capturing at least a portion of said plurality of amplification products using an agent that binds to said detectable label;
- (e) contacting at least a portion of said plurality of amplification products to a guide RNA directed endonuclease and at least one guide RNA that specifically binds to a 5′ end or a 3′ end of at least a subset of said plurality of amplification products, thereby cleaving a 5′ end or a 3′ end of said plurality of amplification products to produce a plurality of cleaved amplification products;
- and
- (f) isolating said plurality of cleaved amplification products, thereby detecting said RNA isoform.
4 more elements
A method of detecting an RNA isoform comprising:
- (a) contacting a plurality of RNA molecules to a oligo(dT) primer and a reverse transcriptase to produce a plurality of cDNA molecules;
- (b) attaching adaptors to a 5′ end and a 3′ end of each of said plurality of cDNA molecules;
- (c) amplifying said plurality of cDNA molecules with a first primer and a second primer that bind to said adaptors to produce a plurality of amplification products;
- (d) contacting at least a portion of said plurality of amplification products to a cleavage deficient guide RNA directed endonuclease and at least one guide RNA that specifically binds to a 5′ end or a 3′ end of at least a subset of said plurality of amplification products;
- (e) capturing at least a portion of said plurality of amplification products using an agent that binds to said cleavage deficient guide RNA directed endonuclease, thereby detecting said RNA isoform.
2 more elements
Dependencies are inferred from the claim text.