Breaking Barriers with M20 Seq: Unveiling the Future of High-Throughput Single-Cell Sequencing

In August 2022, M20 Genomics officially launched M20 Seq a groundbreaking single-cell sequencing technology based on random primers. The introduction of this cutting-edge technology signifies the onset of a new era in single-cell sequencing, extending beyond scientific research towards the prospect of future clinical applications.

Leveraging the M20 Seq technology, M20 Genomics has simultaneously introduced the VITA series - a revolutionary high-throughput single-cell sequencing product line. With the capability for (ultra) high-throughput transcriptome sequencing across an extensive spectrum of samples, species, and full-length transcriptome capture, VITA significantly enhances detection sensitivity and broadly expands the application scope of single-cell technology.

A Glimpse into the Current Possibilities and Challenges of Single-Cell Technology

With its capacity to dissect cellular heterogeneity within complex tissues, single-cell sequencing has surpassed the constraints of traditional bulk transcriptome sequencing. Widely adopted for cellular atlas construction, it is also gaining prominence in clinical research. Furthermore, its potential spans early disease detection, monitoring, and personalized medicine in fields like oncology, immunology, and neuroscience.

Despite achieving technological breakthroughs in high-throughput and multi-omics approaches, challenges persist for single-cell sequencing technology. Issues such as low sensitivity and high costs pose critical barriers to the advancement of single-cell-related research. While most high-throughput single-cell sequencing platforms require a cell viability exceeding 70%, clinical samples commonly undergo freezing or formalin fixation and paraffin embedding (FFPE), leading to extremely low viability. Moreover, many technologies rely on poly(A) tails to capture RNA, limiting read length and providing information only on the 3' end of RNA. Addressing these challenges remains crucial to harness the immense potential of single-cell sequencing capabilities.

M20 Seq: Redefining the Frontiers of Single-Cell Sequencing

Since the debut of the first commercially available single-cell RNA sequencing platform in 2015, numerous technological advancements have been developed by researchers and companies. Despite the rapid progress of single-cell RNA sequencing technologies, limitations persist, especially with the majority of platforms still relying on poly(A) capture.

Through M20 Seq, we initiated a groundbreaking innovation at the technological core, introducing the first random primer-based full-length sequencing technology facilitating high-throughput single-cell sequencing of transcriptomes. With our pioneering technology, we not only elevate potential applications but also refine existing strategies, offering several key advantages:

• Full-Length Transcriptome Sequencing: M20 Seq enables full-length RNA capture, allowing the in-depth exploration of non-coding RNAs, alternative splicing and more thereby introducing unique analytical possibilities and tools.
• Versatile Sample Type Compatibility: M20 Seq accommodates fresh and frozen tissues and effectively addresses previous challenges associated with FFPE samples. Facilitating the analysis of preserved samples, M20 Seq is especially well-suited for centralized laboratory settings.
• High Sensitivity: M20 Seq significantly increases the number of genes detected. Across fresh and frozen tissues, as well as FFPE samples, the median genes per cell can reach up to 5000, 3500, and 2500, respectively.
• Cross-Platform Compatibility: M20 Seq delivers outstanding detection performance on both droplet and microwell platforms.
• Various Species: M20 Seq surpasses technological boundaries, providing comprehensive sequencing capabilities across a diverse range of organisms, including prokaryotic as well as eukaryotic cells - from bacterial to human samples.

Ultra-High Throughput: M20 Seq redefines sequencing capabilities, with its potential to capture up to millions of cells in a single experiment.

The VITA Single-Cell Full-Length Transcriptome Sequencing Platform based on M20 Seq, enables comprehensive single-cell transcriptome studies across various sample types and species, significantly expanding the applications of single-cell technology. Our VITA platform includes the VITApilote kit series, the VITAcruizer single-cell partitioning instrument, and the VITAseer software and data platform (Fig. 1).

Figure 1: VITA Single-Cell Full-Length Transcriptome Sequencing Platform based on M20 Seq

Leveraging the powerful capabilities of our cutting-edge VITA platform, we have attained exceptional results in diverse samples, showcasing the high-performance nature of our innovative product:

We conducted experimental evaluations across diverse sample types, including FFPE, frozen and fresh tissues from human brain, as well as bacterial samples. The results revealed an impressive number of genes detected by our VITA platform (Fig. 2), emphasizing its robust performance across a broad spectrum of sample types.

Figure 2: Number of Genes Detected Across Various Sample Types obtained with VITA Platform

Furthermore, employing our VITA platform led us to uncover the intricate cellular diversity within a frozen liver tissue sample. This approach exposed 13 distinct subpopulations (Fig. 3), showcasing the capabilities of our VITA platform in extracting meaningful insights from complex samples.

Figure 3: Cluster Analysis of VITA Single-Cell Transcriptome Data from a Frozen Human Liver Sample

Analyzing a mixed population of human and mouse cells showcased the remarkable efficacy of our VITA platform, as we achieved notably low doublet rates of less than 1% (Fig. 4), underscoring our platform's precision.

Figure 4: Assessment of Doublet Rate analyzing a Mixed Sample of Mus musculus and Homo sapiens cells with VITA Platform

We additionally assessed the technical reproducibility of our VITA platform by conducting two independent experiments using the same fresh sample of mouse kidney tissue. Comparing UMAP clusters in both replicates revealed consistent patterns (Fig. 5), demonstrating the outstanding reproducibility and stability attainable with our VITA platform.

Figure 5: UMAP Clustering of Two Technical Replicates analyzed with VITA Platform

Unlike technologies relying on poly(A) tails or probes for RNA capture, our VITA platform based on M20 Seq achieves a significant breakthrough, enabling full-length coverage of transcriptomes with the gene body uniformly covered from 5' to 3' (Fig. 6).

Figure 6: mRNA 5'-3' Coverage in VITA Platform Data

M20 Seq not only streamlines the expenses associated with single-cell sequencing but also empowers researchers and clinicians with the opportunity to explore biorepositories by facilitating the sequencing of frozen and FFPE samples. This advancement fosters the application of single-cell technology in pivotal fields such as oncology, reproduction, aging, and immunity.

Beyond our commitment to unlocking unrealized potential in established fields, we're dedicated to leveraging our cutting-edge technology for breakthroughs in potential clinical applications and drug development. With unwavering dedication, we continue to chart the course for progress in life science and technology through our next-generation single-cell technology.