Simultaneous RNA-DNA FISH in Mouse Preimplantation Embryos

Area of Science:

• Cytogenetics
• Molecular Biology
• Genomics

Background:

• Fluorescent in situ hybridization (FISH) is crucial for visualizing and quantifying nucleic acids in cells.
• Existing FISH methods for simultaneous RNA and DNA detection are often compromised by extensive sample processing, leading to damage and time inefficiency.
• Detecting both RNA and DNA in the same sample is challenging due to protocol complexity.

Purpose of the Study:

• To develop a brief protocol for the simultaneous detection of RNA and DNA in a single sample.
• To optimize combined fluorescent in situ hybridization (FISH) for simultaneous RNA and DNA analysis.
• To enable efficient gene expression and genetic aberration analysis in the same cellular context.

Main Methods:

• Developed a streamlined protocol for combined and simultaneous RNA and DNA detection using fluorescent in situ hybridization (FISH).
• Demonstrated the protocol using Xist RNA and centromeric DNA of chromosome 6 in mouse preimplantation embryos.
• Established a method for generating indirect-labeled probes from Bacterial Artificial Chromosomes (BACs).

Main Results:

• Successfully achieved simultaneous detection of Xist RNA and centromeric DNA of chromosome 6 in mouse embryos.
• The protocol significantly reduces processing steps compared to sequential RNA and DNA FISH.
• The method for generating indirect-labeled probes from BACs is effective for combined FISH applications.

Conclusions:

• The developed brief protocol enables efficient simultaneous RNA and DNA detection via FISH.
• This technique minimizes sample damage and processing time, advancing cytogenetics and diagnostics.
• The protocol is adaptable for various RNA and DNA detection combinations in diverse cellular samples.