University of Massachusetts

Carol Villnave Johnson

Jeanne B Lawrence

Yigong Xing

Robert H Singer

Improved methodologies for in-situ hybridization and non-isotopic detection of nucleic acid sequences are provided which offer major increases of resolution, sensitivity, and simplicity unavailable in previously known techniques. The methodology is able to detect even a single-copy of a specific nucleic acid of interest under controlled conditions regardless of whether these are DNA or RNA sequences; or whether the nucleic acid sequence of interest is localized in the chromosomes, nucleus, or cytoplasm of a cell. The methods employ a variety of non-isotopic labels and detection means for rapid and reliable assays. The invention is also provided in kit form for use in the clinical/diagnostic laboratory such that a relatively unskilled person can accurately and reproducibly detect even a single-copy of a specific nucleic acid of interest.

In-situ hybridization of single-copy and multiple-copy nucleic acid sequences

Disclosed are in situ hybridization methods for differentially detecting an RNA and the gene from which it was transcribed, while preserving the spatial relationship of the RNA transcript and the gene. Also disclosed are in situ hybridization methods for simultaneously detecting two alleles of the same gene in a single cell, while differentially detecting RNA transcribed each of the two alleles. Also disclosed are in situ hybridization methods for detecting normal and defective RNA splicing.

Detection of individual gene transcription and splicing

In situ hybridization methods for assessing, determining or observing the RNA produced by transcriptionally active genes. In one embodiment, the methods allow simultaneous observation of the gene and its transcripts in a spatially correlated manner. As the in situ hybridization methods have established their ability to maintain the targeted mRNA at the site of its transcription, it can be determined which genes are being expressed and the level of expression can be quantitated.

Growing community of inventors

Stowe, MA, United States of America

Carol Villnave Johnson