TY - JOUR
T1 - Reporting off-target effects of recombinant engineering using the pORTMAGE system
AU - Sanders, Brittany R.
AU - Townsend, Sydney E.
AU - Ford, Maria L.
AU - Graves, Joseph L
AU - Thomas, Misty
PY - 2023/1/1
Y1 - 2023/1/1
N2 - pORTMAGE recombineering is a simple technique for incorporation of novel point mutations into bacterial genomes that eliminates off-target effects. Here we inserted point mutations into the cusS gene from Escherichia coli, then, using Illumina sequencing, report genetic variants in all mutant strains. Several off-site mutations were found at high frequency. Low frequency mutations also show high heterogeneity. This means that it is essential for studies to report all off-target effects and acknowledge the effect that this may have on resultant phenotypes.
AB - pORTMAGE recombineering is a simple technique for incorporation of novel point mutations into bacterial genomes that eliminates off-target effects. Here we inserted point mutations into the cusS gene from Escherichia coli, then, using Illumina sequencing, report genetic variants in all mutant strains. Several off-site mutations were found at high frequency. Low frequency mutations also show high heterogeneity. This means that it is essential for studies to report all off-target effects and acknowledge the effect that this may have on resultant phenotypes.
KW - Off-target
KW - pORTMAGE
KW - Recombinant engineering
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85143862042&origin=inward
UR - https://www.scopus.com/inward/citedby.uri?partnerID=HzOxMe3b&scp=85143862042&origin=inward
U2 - 10.1016/j.mimet.2022.106627
DO - 10.1016/j.mimet.2022.106627
M3 - Article
C2 - 36436701
SN - 0167-7012
VL - 204
JO - Journal of Microbiological Methods
JF - Journal of Microbiological Methods
M1 - 106627
ER -