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Publication: Mitochondrial Phenotypes Distinguish Pathogenic MFN2 Mutations by Pooled Functional Genomics Screen

In this paper, the researchers presented a generalizable functional genomics platform that combines high-content imaging, machine learning, and CellRaft Array isolation in a new method termed “Raft-Seq”. To demonstrate the efficacy of this platform, this technique was able to distinguish pathogenic point mutations of the mitochondrial regulator MFN2, even when the cellular phenotype is subtle. Raft-Seq enables a new way to perform pooled screening on sets of mutations in biologically relevant cells, with the ability to physically capture any cell with a perturbed phenotype and expand it clonally, directly from the primary screen. Dox-inducible Cas9 cell line was generated using CRISPR HDR. Engineered clones were seeded on a CellRaft Array and isolated into collection plates for further expansion. gRNA libraries were prepared for sequencing.

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