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Updated: Sep 29, 2026
Isolation of Subcellular Ribosome Subpopulations Based on Recombinant Peptide Tag-Specific Location-Restricted Illumination-Enhanced Biotinylation
Published on: July 24, 2026
Protocol for ribosome-protected fragment isolation from rare leukemia cell populations
Christina Mayerhofer1, Ernst Mayerhofer2, Dan Li3
1Center for Cancer Research and Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA, USA; Harvard Stem Cell Institute, Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA; Broad Institute of the Massachusetts Institute of Technology and Harvard University, Cambridge, MA, USA; Department of Pediatric Hematology, Oncology and Stem Cell Transplantation, Children's Hospital, Medical Center and Faculty of Medicine, University of Freiburg, Freiburg, Germany; German Cancer Consortium (DKTK), Partner Site Freiburg, and German Cancer Research Center (DKFZ), Heidelberg, Germany.
Abstract:
Malignant cells under chemotherapy stress alter protein translation, necessitating sensitive methods to profile rare, surviving populations in vivo. We present a protocol for profiling low-input acute myeloid leukemia samples using an optimized ribosome profiling approach. We describe steps for lysing cell inputs, digesting unshielded RNA, enriching ribosome footprints, and gel-based size selection. We then detail procedures for constructing sequencing-ready libraries and using a computational pipeline to align and quantify both protected fragments and matching transcriptomes for reproducible translation analysis. For complete details on the use and execution of this protocol, please refer to Mayerhofer et al.1.
