Summary Plasmid selection and strand replacement synthesis in donor cells during conjugative transfer was examined by a procedure involving electroporation of test plasmid DNA, containing a base pair mismatch, into donor cells prior to mating. Multiple copies of the plasmid were transferred from a ...
Heteroplasmy occurs when wild-type and mutant mitochondrial DNA (mtDNA) molecules co-exist in single cells1. Heteroplasmy levels change dynamically in development, disease and ageing2,3, but it is unclear whether these shifts are caused by selection or d
Cells were transfected with plasmids containing the sgRNA sequences using Lipofectamine 3000 transfection reagent (L3000015, Invitrogen, Waltham, MA, USA). Single clones were screened, amplified, and subsequently employed in downstream experiments. Tumor cell labeling The EO771-PTEN-KO-GFP-Luc cell ...
Accordingly, we present a yeast biocontainment strain that is highly sensitive to fluoride and a complementary set of DNA vectors reliant on fluoride-based selection (Fig. 1). Fig. 1: Engineered fluoride sensitivity augments biocontainment and selection systems. Yeast cells lacking native fluoride ...
Engineering T cell specificity and function at multiple loci can generate more effective cellular therapies, but current manufacturing methods produce heterogenous mixtures of partially engineered cells. Here we develop a one-step process to enrich unlabeled cells containing knock-ins at multiple target lo...
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Summary Plasmid selection and strand replacement synthesis in donor cells during conjugative transfer was examined by a procedure involving electroporation of test plasmid DNA, containing a base pair mismatch, into donor cells prior to mating. Multiple copies of the plasmid were transferred from a donor...
This cassette contains the kanamycin resistance gene flanked by FRT (FLP recognition target) sites, in turn flanked by DNA sequences homologous to the gene targeted for disruption. The gene disruption cassette was transformed into DH5α cells containing the helper plasmid pKD46, which has the λ...
The CRISPR-Cas9 gene editing technique is an innovative tool that enables scientists to make accurate modifications to the DNA of living cells. CRISPR technology has been employed in the creation of genetically engineered crops, the development of novel therapies for genetic disorders, and the ...
As CRISPR effectors like Cas9 increasingly enter clinical trials for therapeutic gene editing, a future for personalized medicine will require efficient methods to protect individuals from the potential of off-target mutations that may also occur at specific sequences in their genomes that are similar ...