Unexpected insertion of carrier DNA sequences into the fission yeast genome during CRISPR-Cas9 mediated gene deletion.
Sophie LongmuirNabihah AkhtarStuart A MacNeillPublished in: BMC research notes (2019)
None of the four microprotein-encoding genes was essential for viability, meiosis or sporulation, and the deletion cells were no more sensitive to a range of cell stressors than wild-type, leaving the function of the proteins unresolved. During CRISPR-Cas9 editing however, a number of strains were isolated in which additional sequences were inserted into the target loci at the Cas9 cut site. Sequencing of the inserts revealed these to be derived from the chum salmon Oncorhynchus keta, the source of the carrier DNA used in the S. pombe transformation.
Keyphrases
- crispr cas
- genome wide
- genome editing
- single cell
- wild type
- circulating tumor
- cell free
- induced apoptosis
- single molecule
- dna methylation
- copy number
- genome wide identification
- cell cycle arrest
- escherichia coli
- cell therapy
- nucleic acid
- stem cells
- endoplasmic reticulum stress
- oxidative stress
- gene expression
- signaling pathway
- bacillus subtilis
- genome wide association