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Barcode sequence could be a good target for developing a species-specific anti-parasite agent based on CRISPR-Cas9

FASEB J. 2020-05-01; 
Yinlai Pan, Yulong Sun, Yilei Wang, Ziping Zhang
Products/Services Used Details Operation
Synthetic sgRNA and crRNA Service … T7 RNA polymerase, rNTP Mix, Murine RNase Inhibitor, T4 DNA ligase, and DNaseI (RNase free) were purchased from New England Biolabs (NEB) (Beijing, China). Cas9‐C‐NLS Nuclease was purchased from GenScript (Nanjing, China) … Get A Quote

摘要

Parasitic infections are a severe issue in many regions of the world. We assume that if a chemical can destroy a DNA barcode sequence, then this chemical could be developed as a species-specific parasiticidal agent. To test this hypothesis, we designed sgRNAs that target the sequences of both a DNA barcode (ITS-2) and a control (5.8S rDNA) in Cryptocaryon irritans. In in vivo tests, we found that exposure to Cas9 mRNA mixed with sgRNAs was able to significantly reduce the hatching rate of tomont and the survival rate of theront. Quantitative Real-time PCR demonstrated that the DNAs of tomont and theront exposed to sgRNAs and Cas9 mRNA were significantly disrupted, no matter whether they were exposed to a single... More

关键词

Cryptocaryon irritans, CRISPR-Cas9, DNA barcode, quantitative real-time PCR, species-specific anti-parasitic drug