Single-cell genomics unlocked
Introduction
What if you could analyze the genome or transcriptome of a complex microbial community or tissue at single-cell resolution–without having to physically isolate individual cells? The requirement to physically isolate cells is one of several barriers limiting the adoption of single-cell sequencing, a technique that promises to grow our understanding of genetic and functional diversity within complex tissues and biological systems.
The impact of single-cell sequencing has been limited by complex, cumbersome, and expensive laboratory workflows. However, there’s good news: A novel library preparation technology from Factorial Biotechnologies is making it possible for researchers everywhere to gain insights from single-cell sequencing at a cost, workflow, and content parity that rivals conventional next-generation sequencing (NGS)—without having to physically isolate cells.
Challenges limiting the adoption of single-cell sequencing
Current library preparation workflows for single-cell sequencing are complex, time-consuming, and have technical limitations that have slowed the adoption of this technology. To achieve single-cell resolution using conventional NGS methods, researchers must isolate individual cells and lyse them prior to library preparation, a process that requires specialized equipment that many laboratories do not have access to. Existing library preparation methods (i.e., plate, droplet, split pool) have highly complex, multi-step workflows that can result in incomplete physical cell isolation, loss of rare or fragile cell populations, and loss of genetic material. Current single-cell library preparation kits can only generate small, targeted panels for DNA and counting applications like ATAC and RNA-seq, and the process of library preparation can take days.
The high costs of single-cell sequencing have been prohibitive as well. Research and core laboratories have limited budgets to conduct the work. Expensive instruments required for physical cell isolation, reagents and enzymes used in library preparation, and the analysis software all add to the high costs of single-cell sequencing. The time-consuming workflow adds to budget strain as researchers spend valuable hours working through tedious protocols that limit productivity.
The untapped potential of single-cell sequencing
With increased access to scalable, high-throughput, and cost-efficient single-cell sequencing, the scientific community stands to gain incredible insights into areas such as:
- Precision oncology: characterize tumors in detail, monitor tumor evolution, and improve patient stratification accordingly.
- Immunology and immuno-oncology: profile the immune repertoire, identify neoantigens, and monitor the tumor microenvironment.
- Cell and gene therapy development: verify gene transfer, ensure sequence integrity, and monitor clonal diversity of cell populations of interest.
- CRISPR quality control and screening: identify on-target edits and off-target edits.
Furthermore, as increased computing power and advances in data science make multi-omic approaches more powerful, there’s a growing need for single-cell library preparation technologies that can be used for whole genome sequencing, RNA-seq, proteomics, and combination analyses.
Factorial’s innovative solution
Factorial’s innovative in-cell library preparation technology provides a solution that will improve access to single-cell sequencing. By turning each cell into a reaction vessel, distinct sequencing libraries can be produced simultaneously within a mixed cell population, eliminating the need to physically isolate cells prior to library preparation. No longer will researchers need to rely on expensive and dedicated equipment to sort cells.
How does it work? Our high-throughput workflow consists of simple buffer exchanges, barcode addition, in-cell amplification, cell lysis, and library clean-up—all performed in a single tube and completed within one day. As a bonus, Factorial provides access to powerful post-sequencing read clustering and analysis software (Figure 1).
Importantly, Factorial’s library preparation technology is compatible with both DNA and RNA samples: depending on your research, our system can be used to build whole genome, whole transcriptome, and target-enriched libraries for single-cell sequencing.
If you’ve been considering using single-cell sequencing to identify and characterize biomarkers, cell populations, or gene expression patterns, learn more about Factorial’s in-cell library preparation technology that is expanding access and accelerating discovery.