CRISPR-Cas9 integrates exogeneous sorting new recombinant DNA in the model tree Populus trichocarpa

One of the major challenges facing researchers is using an efficient homology-directed DNA repair (HDR) to replace the targeted fragment on the genome of tree species with the desired DNA fragment. In the past, researches have been conducted on genetic modifications in mammals, animals, and plants using HDR effector proteins, CtIP and MRE11, to guide double-stranded breaks (DSBs) more precisely. Whereas previously the genome editing using HDR in poplar trees appeared impossible or very complex, here we exhibited that the deficiency of XRCC4, a cofactor for DNA ligase IV that is a key role in nonhomologous end-joining (NHEJ), caused to enhance the HDR efficiency up to 10%, and dramatically decremented polymorphisms. We also could introduce exogenous bleomycin to the poplar genome and generate stable lines resistant to the zeocin antibiotic.

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