Biostatistics · 2024 · 12 citations · 25 references
Life SciencesBiostatisticsGeneticsComputational BiologyGenomicsTranscriptomicsCrispr-cas SystemsSingle-cell AnalysisSingle-cell GenomicsSingle-cell Rna Sequencing AnalysisSingle-cell TranscriptomicsAttenuation BiasBias-variance Tradeoff AnalysisErrors-in-variables RegressionExponential Error ModelsExponential Family DistributionsMeasurement Error ModelsParameter TuningSingle-cell Rna Sequencing
CRISPR genome engineering and single-cell RNA sequencing have accelerated biological discovery. Single-cell CRISPR screens unite these two technologies, linking genetic perturbations in individual cells to changes in gene expression and illuminating regulatory networks underlying diseases. Despite their promise, single-cell CRISPR screens present considerable statistical challenges. We demonstrate through theoretical and real data analyses that a standard method for estimation and inference in single-cell CRISPR screens-"thresholded regression"-exhibits attenuation bias and a bias-variance tradeoff as a function of an intrinsic, challenging-to-select tuning parameter. To overcome these difficulties, we introduce GLM-EIV ("GLM-based errors-in-variables"), a new method for single-cell CRISPR screen analysis. GLM-EIV extends the classical errors-in-variables model to responses and noisy predictors that are exponential family-distributed and potentially impacted by the same set of confounding variables. We develop a computational infrastructure to deploy GLM-EIV across hundreds of processors on clouds (e.g. Microsoft Azure) and high-performance clusters. Leveraging this infrastructure, we apply GLM-EIV to analyze two recent, large-scale, single-cell CRISPR screen datasets, yielding several new insights.
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Pooled CRISPR screening with single-cell transcriptome readout
Paul Datlinger, André F. Rendeiro, Christian Schmidl et al. · Nature Methods · 2017 · 1.1K citations · Full text