Plasmids 101: Knockout/Knock-In Plasmids
Type
Blog Post
Published
Dec. 1, 2016, 3:30 p.m.
...recombinase. If GFP is under control of an endogenous promoter, you can use expression GFP to track cells...tag an endogenous protein with GFP, as seen in blue flame plasmid OCT4-eGFP-PGK-Puro from the Jaenisch lab...In order to both knockout a given gene and insert GFP into the genome, we’d create a plasmid similar to...to the one shown below, where the sequence of GFP is cloned together with the Neomycin resistance (NeoR... 3 of the targeted gene. Upon recombination, the GFP/NeoR cassette is inserted in place of exon 2. Thus...gene is disrupted (knocked out) but the inserted GFP is expressed (knocked in). As seen in the above example...knockout/knock-in targeting vector designed to insert GFP into a given locus.
Conditional knockouts
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