Requesting reagents?

[deleted-103939]

Hi everyone! My project involves targeted gene delivery using chitosan nanoparticles to confer HIV-1 resistance in human cells. The supplies I need for actually developing the agent are a) two plasmids carrying two different genes I need, b) chitosan nanoparticles, and c) a monoclonal antibody targeting the CD4 receptor on cells to be conjugated onto the nanoparticles, along with some other smaller reagents that I acquired/had synthesized easily. The costs for the nanoparticles aren't staggering - $100 or less - and the purified antibodies are only around $60, so I'm not worried about acquiring those.

However, the gene-carrying plasmids I need, despite having been utilized in research several times before, are not readily available commercially, not even on websites like addgene.org. And obtaining them via a gene synthesis service could add around a thousand dollars to my costs. While I have the funding to do so, I'd rather not undertake this amount of spending for something that may later bring up concerns about quality. I know from reviewing scientific literature that the researcher who first cloned one of these plasmids has often gifted the reagent to other researchers for free, and another paper lists an email address to which requests for reagents (including the other plasmid I need) should be directed.

Therefore, I was wondering how to go about politely requesting these reagents in small quantities (around 2 micrograms apiece) from these individuals. I really don't want to cause these individuals too much trouble in providing me access to these reagents, and I'm not unwilling to pay up to a certain amount of money for each. Could anyone give me some advice on this?

Thanks so much!

[donnahardy2]

Hi Everest,

Welcome to Science Buddies!

You should definitely ask for a sample of the plasmid. Many researchers are happy to share samples with other researchers, so I encourage you to start asking for the help that you need. Many researchers will not respond or will say no, but you only need one positive response to be able to continue with your project and save $1000.

If you have the papers of the authors that have used the plasmids that you are interested in, then send an e-mail to the primary author of the paper and politely introduce yourself and explain why you are requesting a sample. If possible, also telephone the researcher's lab and explain your request. This approach will also be helpful in giving you contacts that may be able to provide helpful information in the future. Send as many requests as possible. If there's no response, then send one follow-up e-mail a week later, but then drop the inquiry if you don't hear back.

This is an amazing and very ambitious project, and it sounds like you are doing all of the background work necessary to make this a successful project. Good luck to you, and let us know if you have any questions about doing your experiment.


Donna Hardy

[deleted-103939]

Thanks for the welcome and the advice!

I've started emailing for the plasmids and I've already gotten the positive responses I need! One researcher is providing me with a small sample of one of the plasmids completely free of charge; the other researcher is actually affiliated with a for-profit, commercial institution (that has something of a monopoly over the reagent), so I'll have to pay the lab in order to obtain the reagent. However, the amount I'm required to pay is very small and I saved a great deal of money! So thanks again I really appreciate your input.

And I'll definitely have more questions about my project in the near future

[donnahardy2]

Hi Everest,

Congratulations! Getting the plasmid is a major accomplishment, and means that you will be able to proceed with your project. It sounds like you had to pay a licensing fee, but that’s much better than list price.

Before you continue with this project, I think it would be a good idea to check with your local scientific review committee and make sure your project is approved. Working with plasmids is not specifically prohibited, but since you are working on an HIV vaccine, you need to check at this point. Here’s information this subject from the Science Buddies website (search for SRC), and you can ask your teacher how to contact your local committee. Science projects that don’t meet all safety requirements are disqualified from competition, and you definitely want to avoid this.

https://www.sciencebuddies.org/science- ... ents.shtml

Good luck!

Donna Hardy

[deleted-103939]

Hi again!

Thanks for that information! Here are the aspects of my project that may be the object of concern: a) I actually will be recombining DNA (well, transferring a gene from one plasmid to another), but that's as far as the recombining goes. b) I'll also need to amplify plasmids in bacteria, but not with any intent of "genetically engineering bacteria with multiple antibiotic resistance" - the plasmids will be isolated and purified a few hours after the plasmids have been transformed into the bacteria. c) This step is probably the most concerning for me: After I'm done creating the nanoparticle, I'll have to test it in vitro by using a CD4+ cell line - and an HIV-1 virus (BSL-2 labs are allowed to work with HIV, if I'm not mistaken).

I've evaluated this to be a BSL-2 project, and the lab I'm affiliated with is also a BSL-2 one. So I'll definitely need a form of pre-approval from the SRC; thanks again for notifying me!

Everest

[donnahardy2]

Hi Everest,

Yes, HIV is a BSL-2 pathogen, so if you have access to a lab with this safety level, you should be able to get approval to do your project. I assume that someone will be showing you how to do tissue culture and grow viruses, so there should be no safety concern in doing the experiment if you follow all safety precautions. The experiments you are doing are complicated, and don't always work perfectly the first time, so do plan to start as early as you can in case you have to repeat an experiment. Ideally, you will want to have results in duplicate with similar results so you can make a definitive conclusion.


Donna

[deleted-103939]

Alright, I'll make sure to fill out and submit the appropriate forms (and so will my supervisors)! Thanks for everything