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CRISPR/Cas9-mediated genome editing of Schistosoma mansoni acetylcholinesterase

Here we report successful gene knock-in (KI) in the eggs of Schistosoma mansoni by combining CRISPR/Cas9 with single-stranded oligodeoxynucleotides (ssODNs). We targeted the acetylcholinesterase (AChE) gene of S. mansoni using two synthetic guide RNAs (gRNAs), X5 and X7, respectively. Liver eggs of S. mansoni were exposed to CRISPR-vector containing X5 or X7 by electroporation. Simultaneously, eggs were transfected with a ssODN donor encoding a stop codon in all six frames. Next generation sequencing analysis revealed that CRISPR/Cas9-mediated editing in S. mansoni eggs resulted in Homology-Directed Repair (HDR) when template DNA ssODN provided. Furthermore, soluble egg antigen (SEA) from AChE-modified eggs exhibited markedly reduced AChE activity compared with controls, indicative that programmed Cas9 cleavage mutated the AChE gene. Following injection of modified schistosome eggs into the tail veins of mice, a significant decrease in granuloma size in the lungs of these animals. Notably, an enhanced Th2 response induced by eggs in lung, and splenocytes small intestine-draining mesenteric lymph node cells was also generated in mice injected with X5-KI eggs in different methods. These findings further demonstrate the power and utility of CRISPR/Cas9-based genome editing for undertaking functional genomics studies in schistosomes.

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Policy for DAC EGAC00001001798

Studies are experimental investigations of a particular phenomenon, e.g., case-control studies on a particular trait or cancer research projects reporting matching cancer normal genomes from patients.

Study ID Study Title Study Type
EGAS00001004455 Other

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ID File Type Size Quality Report
Located in
i
EGAF00004838129 fq.gz 39.7 MB Report
EGAF00004838130 fq.gz 55.6 MB Report
EGAF00004838131 fq.gz 12.8 MB Report
EGAF00004838132 fq.gz 19.2 MB Report
EGAF00004838133 fq.gz 20.0 MB Report
EGAF00004838134 fq.gz 33.4 MB Report
EGAF00004838135 fq.gz 19.0 MB Report
EGAF00004838136 fq.gz 32.2 MB Report
EGAF00004838137 fq.gz 21.5 MB Report
EGAF00004838138 fq.gz 36.7 MB Report
EGAF00004838139 fq.gz 20.0 MB Report
EGAF00004838140 fq.gz 33.9 MB Report
EGAF00004838141 fq.gz 13.9 MB Report
EGAF00004838142 fq.gz 23.0 MB Report
EGAF00004838143 fq.gz 17.1 MB Report
EGAF00004838144 fq.gz 29.2 MB Report
EGAF00004838145 fq.gz 15.5 MB Report
EGAF00004838146 fq.gz 22.9 MB Report
EGAF00004838147 fq.gz 18.1 MB Report
EGAF00004838148 fq.gz 26.4 MB Report
EGAF00004838149 fq.gz 13.8 MB Report
EGAF00004838150 fq.gz 20.2 MB Report
EGAF00004838151 fq.gz 14.2 MB Report
EGAF00004838152 fq.gz 21.2 MB Report
EGAF00004838153 fq.gz 12.9 MB Report
EGAF00004838154 fq.gz 18.9 MB Report
EGAF00004838155 fq.gz 21.7 MB Report
EGAF00004838156 fq.gz 35.3 MB Report
EGAF00004838157 fq.gz 16.6 MB Report
EGAF00004838158 fq.gz 24.6 MB Report
EGAF00004838159 fq.gz 19.7 MB Report
EGAF00004838160 fq.gz 32.6 MB Report
EGAF00004838161 fq.gz 16.8 MB Report
EGAF00004838162 fq.gz 24.6 MB Report
EGAF00004838163 fq.gz 14.4 MB Report
EGAF00004838164 fq.gz 21.1 MB Report
EGAF00004838165 fq.gz 20.8 MB Report
EGAF00004838166 fq.gz 34.4 MB Report
EGAF00004838167 fq.gz 18.7 MB Report
EGAF00004838168 fq.gz 31.5 MB Report
EGAF00004838169 fq.gz 17.7 MB Report
EGAF00004838170 fq.gz 29.4 MB Report
EGAF00004838171 fq.gz 15.3 MB Report
EGAF00004838172 fq.gz 22.6 MB Report
44 Files (1.0 GB)