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Plasmid DNA Preparation> | … the mentioned above roles. 149 Here, we aimed to develop the CRISPR/Cas9-mediated knock-in system in poplar with a 150 … Vectors 193 were then extracted using the plasmid midi kit (Qiagen, USA) and confirmed by sanger 194 sequencing (GenScript, Nanjing). 195 … | Get A Quote |
CRISPR-mediated genome editing has become a powerful tool for the genetic modification of biological traits. However, developing an efficient, site-specific, gene knock-in system based on homology-directed DNA repair (HDR) remains a significant challenge in plants, especially in woody species like poplar. Here, we show that simultaneous inhibition of non-homologous end joining (NHEJ) recombination cofactor XRCC4 and overexpression of HDR enhancer factors CtIP and MRE11 can improve HDR efficiency for gene knock-in. Using this approach, the gene was integrated onto the 3' end of the MAP kinase gene to generate a BleoR-MKK2 fusion protein. Based on fully edited nucleotides evaluated by TaqMan real-time PCR, the ... More