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Single-cell ORF perturbation screening of an enriched viral microprotein library in hTERT RPE-1 cells using 10x Genomics Flex fixed-RNA profiling

ABSTRACT: Viral genomes encode large numbers of short open reading frames whose products (microproteins) remain largely uncharacterized. To assay their cellular consequences at scale, we assembled an enriched viral microprotein ORF library in the doxycycline-inducible pANDORA lentiviral vector and delivered it to hTERT RPE-1 cells expressing the pINDUCER20 at low multiplicity of infection (<30% infection rate). The library was split into two sublibraries by insert length ("300mer" and "400mer"). Transduced cells were induced with doxycycline and collected at two time points (24 h and 48 h post-induction), then fixed and profiled by single-cell transcriptomics using the 10x Genomics Single Cell Gene Expression Flex (fixed RNA profiling) assay with 16-plex probe barcoding. ORF identity was read out per cell from a custom probe panel targeting the library inserts, amplified into a dedicated ORF feature library. This dataset comprises paired whole-transcriptome and ORF-assignment measurements for individual cells, enabling the transcriptional consequence of each viral microprotein to be resolved against isogenic controls in the same pool.

ORGANISM(S): Homo sapiens

PROVIDER: GSE344535 | GEO | 2026/08/21

REPOSITORIES: GEO

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Single-cell ORF perturbation screening of an enriched viral microprotein library in hTERT RPE-1 cells using 10x Genomics Flex fixed-RNA profiling

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