The invention relates to the technical field of
plant genomics and
gene editing, in particular to an in-vitro
genome editing technology aiming at wheat and other complex
genome crops, and provides a
plant genome repetitive sequence deletion technology and application thereof. According to the method, specific sgRNA is designed, and CAS9 / CAS12
enzyme is guided to precisely shear
repetitive sequences in a target genome, so that redundant
repetitive sequences in the genome are effectively removed,
sequencing data redundancy is reduced, and the efficiency and precision of genome
information analysis are improved. The method comprises the following steps: firstly, identifying and annotating a
repetitive sequence region by analyzing genome data of crops such as wheat; then, designing an sgRNA sequence aiming at the
repetitive sequence, and optimizing the specificity and amplification efficiency of the sgRNA sequence; then, synthesizing an sgRNA
library by utilizing a high-
throughput DNA synthesis technology, and carrying out an in-vitro shearing reaction together with CAS9 / CAS12
enzyme; and finally, sequencing the sheared genome through a high-
throughput sequencing platform, and analyzing the shearing efficiency and accuracy. The technology not only improves the efficiency of genome simplification and redundant sequence removal, but also can provide powerful data support for
genome research and application of crops such as wheat.