Team:SYSU-CHINA/Results/CyclebowConstruction

Cyclebow construction

Confirmation of Cre’s expression of the STARTER CIRCUIT cell lines

After stable transfecting the STARTER CIRCUIT and antibiotic scanning of Hela and A549 cell lines, we did RT-PCR to confirm the expression of Cre. Samples were collected 24 hours after Doxycycline induction.

From the electrophoresis gel result, it can be seen that both Hela and A549 icre (inducible Cre) stable cell line can express Cre under the induction of Doxycycline. Therefore, our STARTER CIRCUIT cell lines were both confirmed.

Figure 1: RT-PCR and electrophoresis gel results of STARTER CIRCUIT cell lines after 24 hours Dox induction.

Stable cell lines that contains 2 circuits or all 3 circuits of Cyclebow

After obtaining the stable cell lines, we transfected the other circuits in. Due to time limitation, we will transfect COUNTER CIRCUIT and REPROTER CIRCUIT respectively as well as together. As the transfection efficiency was low, and the transfected cells restored population very slow, we were not able to test the complete system before WIKI freeze. Nevertheless, we successully constructed our system.

The following figures are stable transfected cell clones after scanning of:

1. iCre-SmCC Hela (STARTER CIRCUIT+REPORTER CIRCUIT)

2. iCre-SmCC A549 (STARTER CIRCUIT+REPORTER CIRCUIT)

3. iCre-SVE A549 (STARTER CIRCUIT+COUNTER CIRCUIT of pCCNE)

4. iCre-SVE-SmCC A549 (STARTER CIRCUIT+COUNTER CIRCUIT of pCCNE+ REPORTER CIRCUIT)

From the telescope pictures, we can tell that the cells are not yet ready for blocking and inducing.

Figure 2: Stable cell lines that contains 2 circuits or all 3 circuits of Cyclebow