In this review, we discuss the emergence of 4th generation sequencing technologies that preserve the spatial coordinates of RNA and DNA sequences with up to subcellular resolution thus enabling back mapping of sequencing reads to the original histological context. This information is used for example in two current large-scale projects that aim to unravel the function of the brain. Also in cancer research, 4th generation sequencing has the potential to revolutionize the field. Cancer Research UK has named “Mapping the molecular and cellular tumor microenvironment in order to define new targets for therapy and prognosis” one of the grand challenges in tumor biology. We discuss the advantages of sequencing nucleic acids directly in fixed cells over traditional NGS methods, the limitations and challenges that these new methods have to face to become broadly applicable, and the impact that the information generated by the combination of in situ sequencing and NGS methods will have in research and diagnostics. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.