Vol. 12, Issue 7 (2023)

Author(s):
Syam S, Bhavya K, Hirudhaya Ravi, Teenu Paul, JD Khumukcham and Sowmya P

Abstract:
Even with bacterial cells grown under various circumstances, bacterial gene expression is very diverse; bacteria differentiate into subpopulations that may play various roles in the community's survival. Researchers found a distinctive microbial gene regulation mechanism in the quantitative single-cell RNA sequencing using bioinformatics pipelines. Advances in single-cell RNA sequencing (scRNA-seq) have allowed for comprehensive analysis of the bacterial phenotypic community analysis. Prokaryotes' use of scRNA-seq has been limited by their extremely low mRNA quantity, absence of mRNA polyadenylation, and thick cell walls. In the present scenario, as a complementary technique, microSPLiT and PETRIseq use combinatorial indexing-based scRNA-seq procedures to analyse hundreds of bacteria at the same time. Researchers developed MicroSPLiT, a scalable single-cell RNA sequencing technological advancement designed exclusively for bacteria. MicroSPLiT detected a wide range of gene expression levels in over 25,000 single B. subtilis cells, including unusual and unexpected cell states not before observed at the population level. MicroSPLiT is a new approach for analysing single-cell gene expression in complex natural and synthetic microbial communities that has high scalability and resolution. PETRI-seq is a low-cost, high-throughput prokaryotic scRNA-sequencing pipeline. PETRI-seq barcodes transcripts from tens of thousands of cells in a single experiment via in situ combinatorial indexing. PETRI-seq efficiently and accurately collects single-cell transcriptomes of Gram-negative and Gram-positive bacteria. We anticipate that MicroSPLiT and PETRI-seq along with other technologies will be useful in identifying single-cell states and their dynamics in complex microbial diversity.