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Review
. 2015 Dec 14;84(3):612-21.
doi: 10.1128/IAI.01186-15.

A Coming of Age Story: Chlamydia in the Post-Genetic Era

Anna J Hooppaw  1 , Derek J Fisher  2
Affiliations
Review

A Coming of Age Story: Chlamydia in the Post-Genetic Era

Anna J Hooppaw et al. Infect Immun. .

Abstract

Chlamydia spp. are ubiquitous, obligate, intracellular Gram-negative bacterial pathogens that undergo a unique biphasic developmental cycle transitioning between the infectious, extracellular elementary body and the replicative, intracellular reticulate body. The primary Chlamydia species associated with human disease are C. trachomatis, which is the leading cause of both reportable bacterial sexually transmitted infections and preventable blindness, and C. pneumoniae, which infects the respiratory tract and is associated with cardiovascular disease. Collectively, these pathogens are a significant source of morbidity and pose a substantial financial burden on the global economy. Past efforts to elucidate virulence mechanisms of these unique and important pathogens were largely hindered by an absence of genetic methods. Watershed studies in 2011 and 2012 demonstrated that forward and reverse genetic approaches were feasible with Chlamydia and that shuttle vectors could be selected and maintained within the bacterium. While these breakthroughs have led to a steady expansion of the chlamydial genetic tool kit, there are still roads left to be traveled. This minireview provides a synopsis of the currently available genetic methods for Chlamydia along with a comparison to the methods used in other obligate intracellular bacteria. Limitations and advantages of these techniques will be discussed with an eye toward the methods still needed, and how the current state of the art for genetics in obligate intracellular bacteria could direct future technological advances for Chlamydia.

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Figures

FIG 1
FIG 1
Summary of transformation methods available for Chlamydia. The vector backbones drawn in light blue represent regions based on E. coli plasmids (pE. coli). The suicide vectors are used for insertional mutagenesis (intron) or allelic exchange (16/23S rRNA vector). Available markers for selection of transformants and markers that provide a trackable phenotype are listed to the left of the cell diagram.
FIG 2
FIG 2
Chemical mutagenesis methods used for TILLING (A) or forward genetic approaches (B) are shown. The green and red lines in the wells in the TILLING section represent wild-type and mutant amplicons, respectively (see the text). Both methods require whole-genome sequencing to map mutations and may require lateral gene transfer and homologous recombination between the mutant (yellow) and parental (green) strains to isolate mutant alleles. Chemical mutants (and intron-generated mutants) can be complemented using shuttle vectors (Fig. 1).
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