Difference between revisions of "Team:UCLA/Project/Functionalizing Silk"

(Created page with "{{Template:UCLA}} <h2>Functionalizing Silk Fibers</h2> <p> To give functionality to our fibers, we will be genetically fusing various proteins onto our silks. In effect, these ...")
 
Line 4: Line 4:
  
 
<p>
 
<p>
−
To give functionality to our fibers, we will be genetically fusing various proteins onto our silks. In effect, these fusion proteins will have dual function: that of the fused protein as well as that of natural silk protein assembly. Areas like medicine, art, and the fashion industry could immensely benefit from incorporating these dynamic fusion proteins into their current practices.
+
Silk fibers possess the potential to be transformed into functional biomaterials that can be exploited in an array of biomedical applications, from aiding nanoscale drug delivery to simulating medical sutures. However, traditional methods of incorporating functional domains into fibers involve difficult, costly, and time-consuming processes. We propose an in vitro, co-spinning method to quickly and efficiently functionalize silk fibers. In essence, we spin a mixture of wild-type silk dope spiked with a small volume of functional domain. This functional domain which will bind to the native silk proteins when co-spun, thereby incorporating itself into the final synthetic fiber. To ensure proper binding of our functional domain, we created a co-spinning module. This module is a genetic construct consisting of our gene of interest flanked on either side by the N and C terminal domains of Bombyx mori (silkworm silk). When co-spun, the termini on our synthetic protein will bind to the respective termini in the native silk proteins, thereby functionalizing the fiber. Our goal is to develop, optimize and experimentally validate our co-spinning module, and assess its potential as a scalable and powerful tool to manufacture silk fibers with an array of functional capacities.  
 
</p>
 
</p>
  
  
 
<p>
 
<p>
−
To begin with, we will fuse Green Fluorescent Protein (GFP) onto the silk. This will not only be a proof of concept for future experiments, but it will produce a tangible product. Converting this product into, say, a glowing silk t-shirt, is only a couple steps away.
+
Achievements
 +
Successful in vitro co-spin of wild type Bombyx mori silk dope with synthetic protein to produce a fiber that visibly fluoresces!
 +
Designed and sequence-verified our novel co-spinning module with super folded GFP, (sfGFP) sandwiched between the N and C terminal domains of Bombyx mori.
 +
Successful cloning of co-spinning module with E-coli chassis, and successful amplification of part with Polymerase Chain Reaction (PCR). Successful expression and purification of sfGFP protein with N and C termini attached on either side.
 
</p>
 
</p>
  

Revision as of 18:03, 18 September 2015

iGEM UCLA




Functionalizing Silk Fibers

Silk fibers possess the potential to be transformed into functional biomaterials that can be exploited in an array of biomedical applications, from aiding nanoscale drug delivery to simulating medical sutures. However, traditional methods of incorporating functional domains into fibers involve difficult, costly, and time-consuming processes. We propose an in vitro, co-spinning method to quickly and efficiently functionalize silk fibers. In essence, we spin a mixture of wild-type silk dope spiked with a small volume of functional domain. This functional domain which will bind to the native silk proteins when co-spun, thereby incorporating itself into the final synthetic fiber. To ensure proper binding of our functional domain, we created a co-spinning module. This module is a genetic construct consisting of our gene of interest flanked on either side by the N and C terminal domains of Bombyx mori (silkworm silk). When co-spun, the termini on our synthetic protein will bind to the respective termini in the native silk proteins, thereby functionalizing the fiber. Our goal is to develop, optimize and experimentally validate our co-spinning module, and assess its potential as a scalable and powerful tool to manufacture silk fibers with an array of functional capacities.


Achievements Successful in vitro co-spin of wild type Bombyx mori silk dope with synthetic protein to produce a fiber that visibly fluoresces! Designed and sequence-verified our novel co-spinning module with super folded GFP, (sfGFP) sandwiched between the N and C terminal domains of Bombyx mori. Successful cloning of co-spinning module with E-coli chassis, and successful amplification of part with Polymerase Chain Reaction (PCR). Successful expression and purification of sfGFP protein with N and C termini attached on either side.


Next we will attach streptavidin to our silk. Streptavidin is a well-characterized protein useful in molecular biology for its high affinity towards its binding partner, biotin. Both proteins and small molecules can be "biotinylated," allowing them to bind to streptavidin, or in our case, the streptavidin-silk fusion. A simple test we can do to verify this fusion protein is indeed being formed involves biotinylated GFP to act as a visual indicator. After this, we can even try biotinylating enzymes that can function in the body. Silk is ideal in drug delivery as it is sturdy yet biodegradable, and it can act as a scaffold for these enzymes to work in humans.