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Cell Pathology Revealed by Single-Cell Genotype-Phenotype Mapping
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- 1 Intro
- 2 Cell pathology revealed by single cell genotype- phenotype mapping
- 3 The genetic cause of most cancers is known
- 4 Tumor heterogeneity masks mutation co-occurrence
- 5 Droplet microfluidics provides scalability and cost efficiency
- 6 Single cell sequencing resolves tissue heterogeneity
- 7 DNA is the gold standard for genotyping
- 8 Single cell DNA sequencing requires harsh cell lysis
- 9 Proteinase K digestion of chromatin releases DNA and can be heat inactivated
- 10 Multiplexed PCR in droplets selects for the regions of interest
- 11 Uniform target amplification ensures low allele dropout
- 12 Single cell genotyping allows deconvoluting clonal heterogeneity
- 13 To link the genotype to the phenotype we need both
- 14 Single cell immunophenotyping is well established
- 15 Attaching DNA barcodes to antibodies allows for a sequencing readout
- 16 A genotype vector and an antibody count vector is measured for each cell
- 17 To visualize high dimensional data 2D projections are used
- 18 A synthetic cell mixture proves that the method works
- 19 How does a lengthy disease history affect clonal diversity?
- 20 The phenotype maps into a tight compartment
- 21 A patient with atypical blasts, what drives it?
- 22 Divergent differentiation response hints at unchartered variability
- 23 Partnering with Mission Bio helps adopting of method by other labs
- 24 New questions
- 25 HIV remains an incurable disease due to the latent reservoir
- 26 What is special about cells with latent HIV?
- 27 Can we make an assay that exploits this marker?
- 28 The HIV provirus is a perfect biomarker - can DAb-seq exploit it?
- 29 Integrated HIV genomes can reliably be detected in complex samples
- 30 Distinct T cell subcompartments are resolved
- 31 Strong tropism for distinct CD4+ T cell compartments
- 32 Concluding remarks
