• The parts without part number in the tables below are all donated and authorized by the Guangyao Liu research group of South China Agricultural University, rather than from the parts registered by the iGEM teams in previous years.
| Osglub1-sgRNA Expression Cassette | |||
|---|---|---|---|
| Part Numbers | Name | Type | Part Description |
| Backbone (pUC18) | plasmid_backbone | The backbone of pUC18 contains multiple restriction digestion sites that allow digestion on specific sequences, providing a location for insertion into a foreign DNA fragment | |
| 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 |
| snRNA promoter (AtU3b，AtU3d，AtU6-1，AtU6-29) | Coding | It promotes the multi-target editing capability of the CRISPR/Cas9 system in plants | |
| Restriction sites (BamH-Ⅰ, Bsa Ⅰ(1), Bsa Ⅰ(2), Hind Ⅲ) | Coding | It is used to identify and cleave specific DNA sequences for the construction of expression vectors | |
| marker genes (LacZ, ApR) | DNA | As an important marker for recombinant DNA vectors | |
| primers (U-F/gR-R, Pps/Pgs) | primer | The primary role of these primers is to construct and assemble sgRNA expression cassettes in the CRISPR/Cas9 system | |
| Osglb-sgRNA Expression Cassette | |||
|---|---|---|---|
| Part Numbers | Name | Type | Part Description |
| Backbone (pUC18) | plasmid_backbone | The backbone of pUC18 contains multiple restriction digestion sites that allow digestion on specific sequences, providing a location for insertion into a foreign DNA fragment | |
| 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 |
| snRNA promoter (AtU3b，AtU3d，AtU6-1，AtU6-29) | Coding | It promotes the multi-target editing capability of the CRISPR/Cas9 system in plants | |
| Restriction sites (BamH-Ⅰ, Bsa Ⅰ(1), Bsa Ⅰ(2), Hind Ⅲ) | Coding | It is used to identify and cleave specific DNA sequences for the construction of expression vectors | |
| marker genes (LacZ, ApR) | DNA | As an important marker for recombinant DNA vectors | |
| primers (U-F/gR-R, Pps/Pgs) | primer | The primary role of these primers is to construct and assemble sgRNA expression cassettes in the CRISPR/Cas9 system | |
| Cas9 protein Expression Cassette | |||
|---|---|---|---|
| Part Numbers | Name | Type | Part Description |
| Cas9p and its promoter PUbi | Coding | The ubiquitin promoter is active in plant cells and drives expression in plants | |
| marker genes (HPT, Bar) | DNA | Marker genes used to screen successfully transformed cells or tissues | |
| their promoter 2xP35S | Coding | It is used to drive the expression of various exogenous genes | |
| promoter T35S | Coding | Potent and widely used plant promoter | |
| polyadenylation signal sequence aadA(KanR) | signalling | Ensure proper processing and stability of mRNA | |
| replication origin pBR322 | Coding | Versatile and efficient plasmids | |
| enhancer pVS1 replication | Coding | Enhancer sequences of plant viruses | |
| marker gene ccdB | DNA | Genes encoding toxic proteins in Escherichia coli | |
| left and right boundaries (LB, RB) | Coding | The start and end points of a gene or regulatory sequence | |
| left and right arm sequences (GA-L, GA-R) that guide sgRNA expression | Coding | left and right arm sequences (GA-L, GA-R) that guide sgRNA expression is a guide sequence in the CRISPR/Cas system | |
| restriction enzymes (Asc I, Not I) | protein_Domain | Identify and cut the site | |
| digestion sites Bsa I(B-L) | protein_Domain | Restriction enzymes | |
| Bsa I(B-R) | protein_Domain | Restriction enzymes | |