Abstract
Microsporidia constitute a large phylum of eukaryotic obligate intracellular pathogens that can infect a variety of animal hosts. Understanding the biology of microsporidia is severely limited by our current inability to genetically manipulate these parasites. With a growing number of microsporidian genome sequences available and the revolution of successful CRISPR/Cas9 genome editing in virtually every organism tested, the development of DNA transformation techniques would likely lead to rapid and significant progress in understanding microsporidia biology. Here, we outline the challenges in transformation development and review the exciting recent development of DNA transformation and CRISPR/Cas9 genetic manipulation of Cryptosporidium, as well as summarize the successful genetic manipulation strategies in other intracellular eukaryotic parasites, including Plasmodium and Toxoplasma. By providing a list of possible strategies for transforming microsporidia, we hope to facilitate progress in this important area of microbiology and pathogenesis.
Highlights
Microsporidia constitute a large phylum of eukaryotic obligate intracellular pathogens that can infect a variety of animal hosts
There are a number of potential methods for delivery of the DNA, and it is important to consider whether to introduce the DNA into the parasite in an extracellular stage or an intracellular stage inside of the host. Keeping these issues in mind, we describe below the methods that have been successful for other obligate eukaryotic intracellular parasites and consider possible strategies for successful modification of microsporidia
Because Plasmodium genomes do not encode the genes for canonical nonhomologous end joining (NHEJ) for DNA repair, integration into the genome has been achieved with homologous recombination ( Plasmodium does appear to be capable of an alternative NHEJ pathway for DNA repair [21])
Summary
Citation: Reinke AW, Troemel ER (2015) The Development of Genetic Modification Techniques in Intracellular Parasites and Potential Applications to Microsporidia. PLoS Pathog 11(12): e1005283. doi:10.1371/journal.ppat.1005283 Funding: ERT acknowledges support from NIAID R01 AI087528. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist.
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