New basic parts
| Part Numbers | Name | Type | Part Description |
|---|---|---|---|
| BBa_K5167669 | sgRNA of target OsGluB-1 | Coding | The sgRNA acts as a bridge in the CRISPR/Cas9 system, directing the Cas9 protein to specific DNA sequences for cleavage and repair, enabling precise gene editing |
| Bba_K5167670 | sgRNA of target OsGlb | Coding | The sgRNA acts as a bridge in the CRISPR/Cas9 system, directing the Cas9 protein to specific DNA sequences for cleavage and repair, enabling precise gene editing |
Summary
The part BBa_K5167669 is the recognition sequence of sgRNA when CRISPR/Cas9 system targets the rice genome glutelin coding gene OsGluB1.
The part Bba-K51676 is the recognition sequence of sgRNA when targeting the rice genome globulin coding gene OsGlb in the CRISPR/Cas9 system.
CRISPR/Cas9 gene editing causes frameshift mutations in OsGluB1 and OsGlb, and there is an interaction between the mutations of these two genes in regulating the expression of other storage proteins. In other words, when OsGluB1 is mutated alone, it leads to a decrease in the overall lysine content of rice endosperm storage proteins. Only when both genes are mutated simultaneously can the overall lysine content of rice endosperm storage proteins be increased. Because the protein-lysine mainly comes from the glutelin in the storage protein, the effect of these two mutations on the gene expression of the glutelin family is the main factor. The interaction pattern between these two mutations is as follows.
As shown in the figure, when OsGluB1 is mutated alone, although lysine rich glutelin GluA1,2, GluB4,5,7, GluC1,3 and GluB6 with unknown lysine residues are compensatory expressed, the expression of lysine rich glutelin OsGluB2 and GluA3 is synergistically inhibited. More importantly, the original expression levels of GluA3 and GluB2 rank high. OsGluA3 and OsGluB2 are major genes of glutelin family. The synergistic inhibition of OsGluB1 mutations on protein lysine synthesis cannot be compensated for by only compensatory expression of these lysine rich glutelin.
When OsGlb and OsGluB1 mutate simultaneously, it will strengthen the compensatory expression of lysine rich glutelin GluA1,2 and GluB7, and relieve the synergistic inhibition of OsGluB1 mutation on the expression of lysine rich glutelin GluA3. As OsGluA1,2,3 are the major genes of the glutelin family, when their compensatory expression increases (for OsGluA1,2) and their synergistic inhibition of expression (for OsGluA3) is relieved, they can significantly enhance the compensatory synthesis of protein-lysine, compensating for the loss of the expression of lysine rich glutelin major gene OsGluB2 caused by the synergistic inhibition of OsGluB1 mutation.