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US 12,716,092 B2Granted
Method of identifying circular RNA
Assignee10x Genomics, Inc.
InventorJustin Costa
Life of this patent
Estimating the term…
Abstract
The present disclosure relates in some aspects to methods and compositions for analyzing a biological sample, comprising performing rolling circle amplification of a circRNA in situ in a biological sample, thereby generating an RCA product comprising multiple complementary copies of the circRNA and detecting the RCA product in situ at a spatially localized position in the biological sample.
Claims19
A method for analyzing a biological sample, comprising:
- contacting the biological sample with a primer, wherein the primer hybridizes to a target sequence in a target circular RNA (circRNA);
- using a reverse transcriptase to perform rolling circle amplification (RCA) of the circRNA, thereby generating an RCA product comprising multiple complementary copies of the circRNA;
- and
- detecting the RCA product in situ at a spatially localized position in the biological sample;
- wherein the circRNA and the RCA product comprise multiple marker sequences, wherein the combination of the marker sequences identifies the circRNA, and wherein detecting the RCA product comprises detecting the combination of the marker sequences.
2 more elements
A method for analyzing a biological sample, comprising:
- digesting linear RNA in the sample using RNase R;
- contacting the biological sample with a primer, wherein the primer hybridizes to a target sequence in a target circular RNA (circRNA), wherein the target sequence comprises a back-splice junction;
- using a reverse transcriptase having strand displacing activity to perform rolling circle amplification (RCA) of the circRNA, thereby generating an RCA product comprising multiple complementary copies of the circRNA in the biological sample;
- and
- detecting the RCA product in situ at a spatially localized position in the biological sample, wherein detecting the RCA product comprises detecting multiple marker sequences each comprising a complement of a splice junction present in the circRNA.
2 more elements
Dependencies are inferred from the claim text.