A single plasmid based CRISPR interference in Synechocystis 6803 - A proof of concept.
作者： Prithwiraj Kirtania , Barbara Hódi , Ivy Mallick , István Zoltan Vass , Tamás Fehér
DOI: 10.1371/JOURNAL.PONE.0225375
关键词:
摘要: We developed a simple method to apply CRISPR interference by modifying an existing plasmid pCRISPathBrick containing the native S. pyogenes assembly for Synechocystis PCC6803 and named it pCRPB1010. The technique presented here using deadCas9 is easier implement gene silencing in than other techniques as circumvents genome integration segregation steps thereby significantly shortens construction of mutant strains. executed against well characterized photosynthetic genes get clear phenotype validate potential pCRPB1010 work “proof concept”. Targeting non-template strand psbO resulted decreased amount PsbO 50% decrease oxygen evolution rate. template psbA2 psbA3 encoding D1 subunit photosystem II (PSII) single spacer common sequence span two genes, full inhibition both complete abolition protein synthesis, loss photoautotrophic growth arrest. This first report based, completely lesion free episomal expression execution PCC6803.
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