Study On Monocotyledonous Rice And Dicotyledonous Tomentosa Targeting Efficiency Of CRISPR/Cas9 System

CRISPR/Cas9 system is a kind of immune system from bacteria and ancientbacteria, through a RNA mediated, can be completed on the cutting of exogenous DNA.Such characteristics were used to transformed into robust endonuclease enzyme system,compared to the zinc finger endonucleases and talen, it is simple in design, less time consuming and cost low characteristic the once found, has attracted a lot of attention.Since the success of the Cas9 system in 2013, more and more species have been confirmed.But this process, because of understanding the molecular mechanisms of cas9 system deficiencies and cas9 system in some species is high efficiency in editing howeverit leads a lot of off-target phenomenon,this does not cause too much attention.We were constructed for monocotyledonous rice Os XAT gene and dicotyledonous poplar lignin synthesis related enzyme genes cas9 system single target site carrier and three target sites of vector, and then transformed Rice Calli and poplar leaves, resistant transgenic plants, of detection than the results found, in monocot rice gene targeting efficiency was as high as 76.9%, and in dicotyledonous of Populus tomentosa Yang, single target vector was transformed after no effective gene modification is detected, and the carrier of the three target sites in the transformed get 25% positive seedlings.And in Chinese white poplar single target transformation vector, for the promoter of dicotyledonous plants, rice and the same for monocot promoter, but are not detected effective mutation of target gene in Populus tomentosa.(1) through the comparison of Populus tomentosa targeting efficiency in monocot rice and dicotyledonous, found cas9 system for different species, with large differences in gene editing efficiency, in order to well get the mutant plants and need to optimize the cas9 carrier protein, or use a multiple target sites carriers used in miss serious phenomenon of dicotyledonous plants.(2) through comparison of different cas9 protein start sub in Populus tomentosa targeting efficiency, for woody plants of Populus tomentosa, single target CRISPR / cas9 system may in no further optimization, not in the plant tissue expressed successfully.(3) through comparison of different target CRISPR / cas9 system, for woody plantstomentosa, more than a(preferably greater than 3) carrier of target is to complete the target shooting.