Optogenetic is a new method recently, which can accurately control the time and space of biological processes. Phytochrome like proteins and their binding partners found in plant Arabidopsis and Rhodopseudomonas palustris provide a wide range of photoinduced protein binding pairs for possible photogenetic applications, including blue light (CRY2 and CIB), red light (PhyB and PIF) and near-infrared light (BphP1 and PpsR2).
Among these light induced binding pairs, blue light based systems are widely used in cell culture or surface tissue. However, due to the low phototoxicity and large tissue penetrance of red light, PhyB and PIF induced by red light are more suitable as light sensors for photogenetic operation in light sensitive embryonic tissues or deep tissues. Creative BioMart provides a Cre/loxP system stimulated by red light, which can be used to provide accurate spatial and temporal control for gene manipulation in living tissue.
Figure1. Light Activated Cre lox P System (Yen, 2020) The system consists of two parts, optical sensing module and recombinant enzyme module. The light sensing module is a red light induced binding pair, PhyB and PIF6. In Arabidopsis thaliana, PhyB is in its auxiliary phytopigment (PΦB) It binds to PIF6 in the presence of to respond to red light (~ 650 nm). These proteins then act as co transcriptional activators of target genes. PhyB and PIF family members have been successfully designed to create light controlled gene expression systems.
In the recombinase module, Cre was divided into two complementary regions and fused with PhyB and PIF6 respectively. In the presence of red light, the combination of PhyB and PIF6 will activate Cre.
The system allows photoinduced genomic manipulation and constant gene expression after induction. Combined with loxP allele, multiple gene conversion can also be realized through accurate regional light induction for cloning analysis, lineage tracking or cancer inquiry using our system.
| Step | Service Description |
| Expression strategy design | Client provides the target sequence and expression strategy designed referring to customers’ requirement. |
| Plasmid design | Codon optimization Clone the GOI sequence downstream the promoter |
| Plasmid transformation | Transfer the recombinant plasmid into the host |
| Light induction test | The red light is used to stimulate the GOI expression to test the expression efficient. |
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