Correction of copy number induced false positives in CRISPR screens

PLoS Comput Biol. 2018 Jul 19;14(7):e1006279. doi: 10.1371/journal.pcbi.1006279. eCollection 2018 Jul.

Abstract

Cell autonomous cancer dependencies are now routinely identified using CRISPR loss-of-function viability screens. However, a bias exists that makes it difficult to assess the true essentiality of genes located in amplicons, since the entire amplified region can exhibit lethal scores. These false-positive hits can either be discarded from further analysis, which in cancer models can represent a significant number of hits, or methods can be developed to rescue the true-positives within amplified regions. We propose two methods to rescue true positive hits in amplified regions by correcting for this copy number artefact. The Local Drop Out (LDO) method uses the relative lethality scores within genomic regions to assess true essentiality and does not require additional orthogonal data (e.g. copy number value). LDO is meant to be used in screens covering a dense region of the genome (e.g. a whole chromosome or the whole genome). The General Additive Model (GAM) method models the screening data as a function of the known copy number values and removes the systematic effect from the measured lethality. GAM does not require the same density as LDO, but does require prior knowledge of the copy number values. Both methods have been developed with single sample experiments in mind so that the correction can be applied even in smaller screens. Here we demonstrate the efficacy of both methods at removing the copy number effect and rescuing hits from some of the amplified regions. We estimate a 70-80% decrease of false positive hits with either method in regions of high copy number compared to no correction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Artifacts
  • Astrocytoma / genetics
  • Astrocytoma / pathology
  • Brain Neoplasms / genetics
  • Brain Neoplasms / pathology
  • Cell Line, Tumor
  • Cell Proliferation
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • DNA Copy Number Variations / genetics*
  • Datasets as Topic
  • False Positive Reactions
  • Genomics
  • Humans
  • Models, Theoretical
  • Neoplasms / genetics*
  • Neoplasms / pathology
  • Stomach Neoplasms / genetics
  • Stomach Neoplasms / pathology

Associated data

  • figshare/10.6084/m9.figshare.5140057.v3

Grants and funding

This research was funded by Novartis Institutes for BioMedical Research. The funder provided support in the form of salaries for all authors but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.