Systematic discovery of receptor-ligand biology by engineered cell entry and single-cell genomics
Year: 2021
Type: preprint
Abstract: ABSTRACT Cells communicate with each other via receptor-ligand interactions on the cell surface. Here we describe a technology for l e ntiviral-mediated cell e nt ry by e ngineered r eceptor-ligand interaction (ENTER) to decode receptor specificity. Engineered lentiviral particles displaying specific ligands deliver fluorescent proteins into target cells upon cognate receptor-ligand interaction, without genome integration or transgene transcription. We optimize ENTER to decode interactions between T cell receptor (TCR)-MHC peptides, antibody-antigen, and other receptor-ligand pairs. We develop an effective presentation strategy to capture interactions between B cell receptor (BCR) and intracellular antigen epitopes. Single-cell readout of ENTER by RNA sequencing (ENTER-seq) enables multiplexed enumeration of TCR-antigen specificities, clonality, cell type, and cell states of individual T cells. ENTER-seq of patient blood samples after CMV infection reveals the viral epitopes that drive human effector memory T cell differentiation and inter-clonal phenotypic diversity that targets the same epitope. ENTER enables systematic discovery of receptor specificity, linkage to cell fates, and cell-specific delivery of gene or protein payloads. HIGHLIGHTS ENTER displays ligands, deliver cargos, and records receptor specificity. ENTER deorphanizes antigen recognition of TCR and BCR. ENTER-seq maps TCR specificity, clonality and cell state in single cells. ENTER-seq of patient sample decodes antiviral T cell memory.
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W4200363757