Modified CRISPR-based enzymes improve the prospect of inserting entire genes into the genome NatureBiotech
CRISPR-associated transposases enable recombination-independent, multi-kilobase DNA insertions at RNA-programmed genomic locations. However, the utility of type V-K CASTs is hindered by high off-target integration and a transposition mechanism that results in a mixture of desired simple cargo insertions and undesired plasmid cointegrate products. Here we overcome both limitations by engineering new CASTs with improved integration product purity and genome-wide specificity.
enables cut-and-paste DNA insertion with up to 99.4% simple insertion product purity, while retaining robust integration efficiencies on genomic targets. has substantially higher on-target specificity than canonical CASTs, and we identify several novel factors that further regulate targeted and genome-wide integration. Finally, we extend
to other type V-K orthologs and demonstrate the feasibility of -mediated integration in human cell contexts.
France Dernières Nouvelles, France Actualités
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