Application of CRISPR/Cas9 Technology in Obtaining Double Mutant Materials of Arabidopsis thaliana

doi:10.11924/j.issn.1000-6850.casb17110102

Abstract

Abstract: To study the gene function of sb3 and sb6 containing two MATH domains, we need to obtain the double mutants of these two closely linked genes. CRISPR/Cas9 gene editing technology was used to create special double knockout mutants of sb3 and sb6 genes in Arabidopsis thaliana Col-0., 10 T0 transgenic plants were obtained by PCR and Singer Sequencing. Among them, 5 transgenic plants had no editing in target sequence. In the other five transgenic plants, the target sequence of sb3 gene was edited, but the target sequence of sb6 gene was edited only in 2 transgenic plants. So far, the double-knockout mutants of sb3 and sb6 genes are successfully obtained, belonging to nucleotide deletion mutants. The two double mutants lay a good foundation for further research on the function of dual MATH-domain genes in Arabidopsis thaliana.

References

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