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Flura-Seq Identifies In Situ Transcriptomes of Micrometastases

ABSTRACT: Understanding the biology of rare cell populations in the context of their microenvironment requires an accurate analysis of the transcriptomes of these cells as expressed in situ. We developed fluorouracil-tagged RNA sequencing (Flura-seq) to characterize the transcriptomes of small cell subpopulations from a whole organ in model systems. The method utilizes cytosine deaminase (CD), which converts the non-natural pyrimidine base fluorocytosine to fluorouracil. Expression of S. cerevisiae CD in cells of interest and exposure to fluorocytosine generates fluorouracil, which is metabolically incorporated into newly synthesized RNAs. The fluorouracil-tagged RNAs can then be immunopurified and sequenced. We applied Flura-seq to define the transcriptome of human breast cancer xenografts, representing as few as 0.003% of host organ cell population during the early stages of metastatic colonization of mouse lungs. The robustness, simplicity and lack of toxicity of Flura-seq make this tool broadly applicable to many studies in developmental, regenerative, and cancer biology.

ORGANISM(S): Mus musculus Homo sapiens

PROVIDER: GSE93605 | GEO | 2019/01/01

REPOSITORIES: GEO

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Flura-Seq Identifies In Situ Transcriptomes of Micrometastases

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