Elucidating the wiring diagram of the human cell is a central goal of the postgenomic era. We combined genome engineering, confocal live-cell imaging, mass spectrometry, and data science to systematically map the localization and interactions of human proteins. Our approach provides a data-driven description of the molecular and spatial networks that organize the proteome. Unsupervised clustering of these networks delineates functional communities that facilitate biological discovery. We found that remarkably precise functional information can be derived from protein localization patterns, which often contain enough information to identify molecular interactions, and that RNA binding proteins form a specific subgroup defined by unique interaction and localization properties. Paired with a fully interactive website (opencell.czbiohub.org), our work constitutes a resource for the quantitative cartography of human cellular organization.

MLL gene rearrangements were detected with a single probe and a single restriction-enzyme digest in all DNA samples from patients with the common 11q23 translocations as well as in 16 patients or cell lines with other 11q23 anomalies. The ability to detect an MLL gene rearrangement rapidly and reliably, especially in patients with limited material for cytogenetic analysis, should make it possible to identify patients who have a poor prognosis and therefore require aggressive chemotherapy or marrow transplantation.

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Two genes have been implicated in leukemias of patients with ab l of chromosome 3, band q26: EVII, which can be activated over long dances by chromosomal rearrangements involving 3q26, and EAP, a ribosomal gene that fuses withAMLI in a therapy-related myelodysplasia patient with a t(3;21)(q26.2;q22). AMLI was identified by its involvement in the t(8;21)(q22;q22) of acute myelod leukemia. Here we report the consistent identification of fusion transcripts between AMLI and EAP or between AMLI and previously unidentified sequences that we named MDSI (MDSassociated sequences) in the leukemic cells of four patients with therapy-related myelodysplala/acute myeloid leukemia and in one patient with chronic myelogenous leukemia in blast crisis, all of whom had a t(3;21). In addition, we have identified a third chimeric transcript, AMLI /EVI, in one ofthe therapyrelated acute myeloid leukemia patients.-field gel electrophoresis established the order of the genes as EAP, the most telomeric, and EVII, the most centromeric, gene. The results indicate that translocations could involve multiple genes and affect gene expression over long distances.The molecular analysis of recurring chromosomal translocations in leukemias has led to the identification of protooncogenes located at the translocation breakpoint that are activated either by altered expression or by gene fusion. One of the most common translocations in acute myeloid leukemia is the t(8;21)(q22;q22), which has recently been shown to involve the AMLI gene at 21q22 (1) fused to the ETO gene at 8q22 (2, 3). AMLI is identical' to the murine transcription factor pebp2a and the DNA-binding subunit of the human transcription core factor CBF (1, 4). The human and murine AMLi polypeptides are 99% homologous in their first 242 residues, but they differ in overall size. pebp2a encodes a predicted polypeptide of 452 residues containing the DNAbinding and dimerization region encoded by the Drosophila melanogaster runt (run) homology segment at the N terminus (2, 5), as well as a region rich in serine, threonine, and proline, suggestive of a transcription activation domain at the C terminus (4). The sequence ofthe human AMLI cDNA is 250 residues shorter than that of pebp2a and lacks the serine-, threonine-, and proline-rich region. It probably represents a spliced isoform of the mRNA consisting mostly of the run homology segment.Chromosome 21, band q22, is also involved in the t(3;21)(q26.2;q22) in therapy-related acute myeloid leukemia/myelodysplasia (t-AML/t-MDS) or chronic myelogenous leukemia in blast crisis (CML-BC) (6, 7). Recent studies have shown that this translocation involves the AMLJ gene and a gene at 3q26, which is EAP (8, 9). EAP codes for a small (129 amino acids) ribosomal protein, L22, and belongs to a large family of genes. Although EAP is highly conserved and abundantly expressed in all tissues (10, 11) and in hematopoietic cell lines (9), it is not known whether the allele at 3q26 is the one that is expressed. In t(3;21), the translocation results in th...

The advent of image-activated cell sorting and imaging-based cell picking has advanced our knowledge and exploitation of biological systems in the last decade. Unfortunately, they generally rely on fluorescent labeling for cellular phenotyping, an indirect measure of the molecular landscape in the cell, which has critical limitations. Here we demonstrate Raman image-activated cell sorting by directly probing chemically specific intracellular molecular vibrations via ultrafast multicolor stimulated Raman scattering (SRS) microscopy for cellular phenotyping. Specifically, the technology enables real-time SRS-image-based sorting of single live cells with a throughput of up to~100 events per second without the need for fluorescent labeling. To show the broad utility of the technology, we show its applicability to diverse cell types and sizes. The technology is highly versatile and holds promise for numerous applications that are previously difficult or undesirable with fluorescence-based technologies.