the MYSIS Project

View As One PageView As Slideshow     The MYS!S Project: BioBrick Optimizer Abstract MYS!S is a stand alone software tool designed to assist an “iGEMer” when designing a new BioBrick part or modifying an existing one. MYS!S handles several problems that a synthetic biologist might encounter in the lab that can be solved with site directed mutagenesis. The utilities that MYS!S supports to modify DNA sequences on paper, can be “actualized” in the lab using site directed mutagenesis to change physical DNA “stored” in a backbone. MYS!S will determine the most efficient modified sequence by optimizing for the least number of rounds of site directed mutagenesis. Once the changes are finalized primers are designed. In addition to primer design, MYS!S provides lab protocols for both site directed mutagenesis and transformation into competent cells. Currently the lab protocols are supplied as a BioCoder compliant C++ file, that can then be compiled with the BioCoder library.

Operations
I. DNA Optimization First, it performs DNA codon optimization based on the target organism by removing rare tRNAs and replacing them with more prevalent tRNAs. (We noticed that many teams decided to utilize sequences from a variety of organisms such as firefly luciferase to ultimately be transformed into an experimentally “stable” organism.)   II. Processing Secondly, the program checks the coding sequence for any restriction sites that are not allowed by the format specified by the user. After checking the sequence, MYS!S removes the restriction sites while ensuring that the modified sequence is still codon optimized. In addition, it was pointed out that frequently a synthetic biologist would like to change specific amino acids in the DNA coding sequence. MYS!S decides what difference in nucleotide sequence are required to best make the amino acid change.   III. Restriction site analysis Alternatively, an iGEMer might also want to add restriction sites to a DNA sequence in order to remove a particularly useful sequence. The program can handle this by either allowing the user to specify where the restriction site should be or by choosing what restriction site is desired and how sites need to be added to the sequence.

How Mys!S Does it
The MYS!S Environment One of the main ideas behind MYS!S was to make synthetic biology more accessible to a wider “spectrum” of people. We want the program to provide an environment that enables the user to understand and explore the changes being made to the DNA in a more intuitive manner. Custimization In addition, MYS!S is designed to be highly customizable by providing the user with the ability to add and modify components such as an organism’s codon usage table and new assembly standards. The ultimate goal is to create a program that can be customized by the user for their specific work and the lab protocols they are comfortable with.

MYSIS: An Introduction

Walk through One of the main ideas behind MYS!S was to make synthetic biology more accessible to a wider “spectrum” of people. We want the program to provide an environment that enables the user to understand and explore the changes being made to the DNA in a more intuitive manner. In addition, MYS!S is designed to be highly customizable by providing the user with the ability to add and modify components such as an organism’s codon usage table and new assembly standards. The ultimate goal is to create a program that can be customized by the user for their specific work and the lab protocols they are comfortable with. To showcase the capabilities of MYS!S we would like to walk you through an analysis of a current BioBrick in the registry. For this example we are going to use part BBa_K191006 which is the protein coding sequence for LovTAP. LovTAP was used by both of our laboratory projects Etch-a-Sketch and Full Adder. One of the issues with LovTAP is that it contains restriction sites not allowed by some BioBrick assembly standards. Lets say we want to transform the LovTAP coding region into e-coli After opening MYS!S, navigate to the screen to manage components. To do this go to the MYS!S menu and click “Manage Components”.   It will open up a screen that will allow the user to manage which organisms and assembly standards the program handles. From this screen the user can add organism codon tables, modify existing ones, and delete those not needed anymore. The same functionality applies to standards. The user can specify the prefix, suffix, and the restriction sequences that the nucleotide sequence should not contain.         Now we are going to exit the manage components screen and open a new assembly standard check. You can find the assembly standard check option by going under the file menu then click “New” then click “Assembly Standard Check”.     Next, we need to enter all the information required to perform an assembly standard check. The fields that have to be filled in are the name field, the organism, the standard, and the plasmid. Obviously the user also needs a nucleotide sequence to analyze. This sequence needs to be entered in the original DNA sequence text area. Note MYS!S requires the nucleotide sequence to start with ATG and be in frame.     After these fields are completed the user can hit the go button to perform an assembly standard check. The original DNA sequence and the modified sequence are displayed top and bottom. All proposed changes in the modified sequence are colored green the corresponding nucleotides in the unmodified sequence are colored purple. The primers needed to transform the original Lovtap sequence to the assembly standard acceptable Lovtap are shown in alignment with the 5’ and 3’ ends labeled.