A laboratory-evolved CRISPR-associated transposase adapts to human cells
The transposses (casts) associated with CRISPR are an attractive candidate for genome editing applications, because they allow the insertion of large DNA cargoes without creating double -strand ruptures. However, the casting systems have shown limited activity in human cells. In an article published in ScienceWitte et al. Apply the continuous evolution assisted by phages (rhythm) to direct the rapid evolution of the new distribution variants, by acquiring a casting system capable of effectively integrating cargoes of gene size in human cells.
Rhythm iterative cycles have given an evolved TNSB – a component of the type in the event of Cast transposition machinery – with the integration efficiency in Hek cells more than 200 times higher than that of the wild type. The evolved TNSB contained ten mutations improving the activity covering several areas, which suggests that the pace has optimized various features to improve the performance of TNSB and that obtaining such a variant by rational protein engineering would have been unlikely. In particular, the evolved TNSB does not require supplementation with the CLPX accessory protein, a cytotoxic factor previously used to increase the effectiveness of funding of funds.
