Single-cell RNA sequencing has recently made strides that make it a powerful tool for detecting cellular heterogeneity in a variety of heterogeneous tissues or cell samples.
In this study, we are using droplet microfluidics, one of the most promising high-throughput methods for quickly and effectively capturing and processing huge numbers of individual cells for whole-transcriptome or genomic analysis. However, we will encapsulate individual Yersinia pseudotuberculosis bacterial cells under T3SS in semi-permeable hydrogel microcapsules (SPCs) for single-cell analysis rather than employing droplets. Following encapsulation, the SPCs will house molecular biology procedures such as cell lysis, mRNA capture, cDNA synthesis and PCR, and barcoding employing barcoded oligos of the bacteria cell.
Finally, we will break the SPCs, demultiplex the cells, and utilize computational techniques to cluster the cells based on the different genes that are present in each cell. By doing so, we will be able to recognize the heterogeneity and sub-population associated with T3SS in Yersinia pseudotuberculosis thereby shedding light on bacterial infections.