Concurrent analysis of genome and transcriptome in one single cell.

Area of Science:

• Genomics
• Molecular Biology
• Biotechnology

Background:

• Current single-cell analysis methods often lack a simple, integrated approach for simultaneous whole-genome and whole-transcriptome profiling.
• Analyzing both DNA and RNA from the same cell is crucial for understanding gene regulation and somatic mutations.

Approach:

• Developed a three-step method for physical separation of cytoplasmic RNA from the nucleus of a single, immobilized cell.
• The process involves cell immobilization, hypotonic lysis, and separation of the cytosolic and nuclear fractions.
• This technique allows for the simultaneous extraction of DNA and RNA from individual cells.

Key Points:

• Extracted DNA and RNA are suitable for downstream sequencing applications.
• Transcriptome and genome sequencing coverage is comparable to methods without prior separation.
• The separation process does not introduce significant bias in mutational load or spectra.

Conclusions:

• This updated method provides a simple tool for simultaneous, in-depth analysis of the genome and transcriptome from single cells.
• Enables comprehensive studies on the interplay between somatic mutations and gene expression regulation.
• Facilitates a deeper understanding of cellular processes at the single-cell level.