Cookies on this website
We use cookies to ensure that we give you the best experience on our website. If you click 'Continue' we'll assume that you are happy to receive all cookies and you won't see this message again. Click 'Find out more' for information on how to change your cookie settings.

Insertional mutagenesis in a haploid background can disrupt gene function. We extend our earlier work by using a retroviral gene-trap vector to generate insertions in >98% of the genes expressed in a human cancer cell line that is haploid for all but one of its chromosomes. We apply phenotypic interrogation via tag sequencing (PhITSeq) to examine millions of mutant alleles through selection and parallel sequencing. Analysis of pools of cells, rather than individual clones enables rapid assessment of the spectrum of genes involved in the phenotypes under study. This facilitates comparative screens as illustrated here for the family of cytolethal distending toxins (CDTs). CDTs are virulence factors secreted by a variety of pathogenic Gram-negative bacteria responsible for tissue damage at distinct anatomical sites. We identify 743 mutations distributed over 12 human genes important for intoxication by four different CDTs. Although related CDTs may share host factors, they also exploit unique host factors to yield a profile characteristic for each CDT.

Original publication

DOI

10.1038/nbt.1857

Type

Journal article

Journal

Nat Biotechnol

Publication Date

29/05/2011

Volume

29

Pages

542 - 546

Keywords

Alleles, Bacterial Proteins, Bacterial Toxins, Cell Line, Tumor, Chromosomes, Human, Cloning, Molecular, Genetic Association Studies, Genome, Human, Gram-Negative Bacteria, Haploidy, High-Throughput Nucleotide Sequencing, Humans, Mutagenesis, Insertional, Mutation, Phenotype, RNA Interference, Sequence Tagged Sites, Virulence Factors