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Single-stranded binding proteins and helicase enhance the activity of prokaryotic argonautes in vitro.

PLoS ONE. 2018; 
Hunt Eric A,Evans Thomas C,Tanner Nath
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Recombinant Proteins .... sequence with amino-terminal hexahisti- dine tag from GenScript in plasmid pET28c and transformed.... with amino-terminal hexahistidine tag from GenScript in plasmid pET29a(+) and transformed into NEB Turbo Get A Quote

摘要

Prokaryotic argonautes are a unique class of nucleic acid-guided endonucleases putatively involved in cellular defense against foreign genetic elements. While their eukaryotic homologs and Cas protein counterparts require single-stranded RNAs as guides, some prokaryotic argonautes are able to utilize short single-stranded DNAs as guides for sequence-specific endonuclease activity. Many complications currently prevent the use of prokaryotic argonautes for in vivo gene-editing applications; however, they do exhibit potential as a new class of in vitro molecular tools if certain challenges can be overcome, specifically the limitations on substrate accessibility which leads to unequal levels of activity acros... More

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