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GENERATE KO WITH ZFNs

ZFNs are attractive tools which allow edit genomes and are significantly empowered when combined with the efficiency of viral vector delivery methods. Indeed transfection techniques do not allow to fully use the potential of these genome editing tools, for in vitro, in vivo and transgenesis applications. Including a ZFN in an episomal lentiviral vector allows highly enhancing the efficiency of this tool as illustrated below with ZFNs which target GFP sequence (figure). These ZFNs have been assayed on a Hela cell line which contains the GFP coding sequence in its genome. The results demonstrate that targeting GFP sequence with Lenti-ONE™ Epi ZFN is much more effective than with transfected ZFN plasmids: GFP-ZFN can knock out up to 60% of GFP expression when delivered with a Lenti-ONE™ Epi vs only 2% when delivered through plasmid transfection.

 


 

ZFNs: PLASMIDS TRANSFECTIONS vs Lenti-ONE™ Epi TRANSDUCTIONS

PLASMIDS ZFNs TRANSFECTIONS V.S. Lenti-ONE™ Epi ZFNs TRANSDUCTIONS

Treatment of a Hela cell line containing the GFP coding sequence in its genome with (i) plasmids (D1: 10ng, D2: 100ng) or (ii) Lenti-ONE™ Epi (100ng of p24) ZFN3 and ZFN4 which target GFP coding sequence.

 GFP expression shows up to a 60% descrease when ZFNs are delivered with a Lenti-ONE™ Epi vs only 2% when delivered through plasmid transfection

 


 

Consequently, Lenti-ONE™ Epi ZFN are tools of choice to generate KO in vitro, in vivo or in transgenesis. If you want to vectorize your own ZFNs, contact our On-Demand Services.

To generated KO cell lines with this tool, see to the TechNote How to transduced cells with Lenti-ONE™ Epi. Furthermore, if you need clonal KO cell lines, refer to the TechNote Generate clonal cell lines.