COLLABORATION

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I.Collaboration with NJU_CHINA(CLICK)

NJU_CHINA helped us by constructing one of our TAL effectors (TALE1) using Golden Gate technique. They also edited the original vector pTAL1 provided by the Kit to add EcoRI, SpeI and PstI cutting cite, thus ensuring the engineered TAL effector can meet the RCF10 bio-brick standard. This new vector was then used when we construct the other two TAL effectors ourselves. We really appreciate their assistance in our experiment.

In return, we performed nanoparticle tracking analysis (NTA) for NJU_CHINA to help them to have a more precise determination of the quantity and size of secreted exosomes.

The use of Nanosight enabled quantification and size determination of the EV, as nanoparticles can be automatically tracked and sized based on Brownian motion and the diffusion coefficient. Because exosomes are more homogenous and generally smaller than most EVs with a diameter size ranging from 40 to 120 nm, the percentage of nanoparticles whose size ranges from 40 to 120 nm could be a good indicator of total amount of exosomes. The shift of peak of size distribution of EV from 170 nm to 120 nm and the significant raise of the peak indicated the increase of relative level of exosomes in secreted EVs after overexpression nSMase2 in HEK293 cells.

The result proved the feasible and effective improvement of manufacturing exosomes by overexpressing nSMase2.

Figure 1. Characterization of secreted exosomes after overexpression of nSMase2 in HEK293 cells. (A and B) Representative screen shots of the NTA videos for EV from HEK293 cells under normal condition or after transfection with nSMase2 plasmid. (C-F) Size and intensity of EV from HEK293 cells under normal condition or after transfection with nSMase2 plasmid. (G) Concentration of different particle sizes of exosomes with (red line) or without (blue line) transfection with nSMase2 plasmid. The peak of size distribution of EV shifted from 170 nm to 120 nm after transfection with nsMase2 plasmid, indicating the increase in quantity of secreted exosomes.

You can goto NJU_CHINA Collaborations Pages to affirm our contributions.


NUDT_CHINA	NJU_China

II.Collaboration with USTC-software(CLICK)

This year, we helped USTC-software by sending our part to them, which we submitted last year numbered for BBa_k1532006.

They asked us for help because they needed the plasmid luxR numbered for C0062 in their project, but what they obtained in the kit plate was proved to be wrong by sequencing. As luck would have it, we submitted it as one of our bricks last year, so we sent it to them as soon as possible to ensure their project runs smoothly.

You can goto USTC-software’s Wiki to affirm our contributions.


NUDT_CHINA	USTC-software

III.Collaboration with NEFU_China(CLICK)

Wet Lab: Sent a plasmid vector to NEFU_China

This year, we helped NEFU_China by sending a plasmid vector to them.

We got in touch with NEFU_China in the 2015 CiCC (China iGEMer Committee). When we exchanged ideas with others, they expressed the difficulty they faced at that time that they were lack of a pivotal plasmid vector which is too hard to buy. We were pleased to offer it to them as we have stored it in our plasmid library. We can obtain a huge sense of satisfaction by helping our friends get over the trouble.

You can goto NEFU_China’s Attribution Page to affirm our contributions.

We used and improved NEFU_China’s “Flight iGEM”

This year, we helped NEFU_China to improved their “Flight iGEM” platform which aims to build a friendly website to make the wiki development easier.

We got touch with NEFU_China in the 2015 CiCC (China iGEMer Committee). Then, we participated in the “Flight iGEM” beta test. Below is our contributions to “Flight iGEM”:

1. Improve the Human-Computer Interaction (HCI) for the “Flight iGEM” Magic function.

2. Figure out the BUG in the login bar which is significant because the 2015 wiki new rules forbad the revise of login bar.

3. Contact with the igem.org to make sure the feasibility of cleariGEM template.

We are the users of "Flight iGEM"

You can goto NEFU_China’s Software page to affirm our contributions and more details about the “Flight iGEM”.