Understanding full-length single-cell RNA-seq of immune cells in cardiovascular diseases

Discover how full-length isoform sequencing enables you to robustly and cost effectively identify new isoforms at scale and study isoform dynamics in single-cell populations. The MAS-Seq for 10x Single Cell 3’ kit – and its successor, the Kinnex single-cell RNA kit – concatenate isoforms for highly accurate long-read sequencing (HiFi sequencing). This approach increases throughput and reduces costs by giving you a 16-fold throughput increase compared to un-concatenated libraries, without losing accuracy or full-length isoform information.

Anastasiya Grinko, MSc, applied the MAS-Seq kit to profile the single-cell isoform landscape of monocyte-derived macrophages after myocardial infarction. This approach captured thousands of distinct isoforms, making it possible to see cell-type-specific isoform utilization in monocytes and macrophage subtypes and to unveil temporal dynamics of isoform expression.

Watch this webinar to:
• Understand the importance of elucidating isoforms in single-cell transcriptomics.
• Learn how to analyze MAS-Seq and Kinnex single-cell data with the Single-cell Iso-Seq workflow in SMRT Link.
• See how the throughput of the MAS-Seq single-cell kit revealed differentially expressed isoforms across cell types in heart tissue after myocardial infarction.

00:00 Introduction to the Kinnex single-cell RNA kit
01:50 Example of single cell isoform sequencing using HiFi
03:04 The MAS-Seq method for increasing throughput
04:29 Throughput and compatibility of the Kinnex single cell RNA kit
05:30 Public dataset for Kinnex single cell RNA
06:31 Speaker Anastasia Grinko
08:03 Why use the MAS-Seq method and Kinnex kits?
09:32 Applying the MAS-Seq method to cardiac immune cells
10:08 Quality metrics and data analysis
11:59 Processing and pipeline
12:28 Comparing short and long reads
13:49 Gene and isoform distribution
14:14 Comparison of short-read vs long-read single cell data
14:52 Cell type-specific isoform usage
15:35 SQANTI3/pigeon for isoform classification
22:54 Differential splicing analysis
33:24 Conclusion and final remarks

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