PT  - JOURNAL ARTICLE
AU  - Madeline E. McCarthy
AU  - William B. Dodd
AU  - Xiaoming Lu
AU  - Nishi D. Patel
AU  - Charlotte V. Haskell
AU  - Hugo Sanabria
AU  - Mark A. Blenner
AU  - Marc R. Birtwistle
TI  - A Theory for High-Throughput Genetic Interaction Screening
AID  - 10.1101/2022.10.05.510977
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.10.05.510977
4099  - http://biorxiv.org/content/early/2022/10/05/2022.10.05.510977.short
4100  - http://biorxiv.org/content/early/2022/10/05/2022.10.05.510977.full
AB  - Systematic, genome-scale genetic screens have been instrumental for elucidating genotype-phenotype relationships, but approaches for probing genetic interactions have been limited to at most ∼100 pre-selected gene combinations in mammalian cells. Here, we introduce a theory for high-throughput genetic interaction screens. The theory extends our recently developed Multiplexing using Spectral Imaging and Combinatorics (MuSIC) approach to propose ∼105 spectrally unique, genetically-encoded MuSIC barcodes from 18 currently available fluorescent proteins. Simulation studies based on constraints imposed by spectral flow cytometry equipment suggest that genetic interaction screens at the human genome-scale may be possible if MuSIC barcodes can be paired to guide RNAs. While experimental testing of this theory awaits, it offers transformative potential for genetic perturbation technology and knowledge of genetic function. More broadly, the availability of a genome-scale spectral barcode library for non-destructive identification of single-cells could find more widespread applications such as traditional genetic screening and high-dimensional lineage tracing.Competing Interest StatementThe authors have declared no competing interest.