CRISPR-based Genome Editing Systems For Editing in E. coli
Type
Blog Post
Published
April 13, 2021, 1:15 p.m.
...sequence in pEcgRNA. Next both plasmids are transformed in E. coli with the donor DNA for editing to take place...beneficial for plasmid clearance.
It is more convenient to update the spacer(20nt) on pEcgRNA plasmid as only....
pCutamp cures the plasmids by targeting the ampR promoter in all three plasmids. pCutamp is then removed... 13 plasmids with 14 rounds of transformation in 30 days.
Figure 2: (A) MUCICAT plasmids for ... original pCas/pTargetF system consists of one plasmid that expresses the S. pyogenes Cas9, a temperature-sensitive...the pTargetF construct and then introduce both plasmids into the cell. The edit in the chromosome can ...the pMB1 replicon, resulting in cutting of the plasmid.
At this point, subsequent introductions of pTargetF...