iGEM 2020 — Team Hong_Kong_HKU

[![](https://static.igem.org/mediawiki/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:Hong Kong HKU/Notebook

# Notebook

Document the dates you worked on your project. This should be a detailed account of the work done each day for your project.

### What should this page have?

* Chronological notes of what your team is doing.
* Brief descriptions of daily important events.
* Pictures of your progress.
* Mention who participated in what task.

### Inspiration

You can see what others teams have done to organize their notes:

* [2018 Munich](https://2018.igem.org/Team%3AMunich/Notebook)
* [2014 ATOMS-Turkiye](https://2014.igem.org/Team%3AATOMS-Turkiye/Notebook)
* [2014 Tec Monterrey](https://2014.igem.org/Team%3ATec-Monterrey/ITESM14_project.html#tab_notebook)
* [2014 Kyoto](https://2014.igem.org/Team%3AKyoto/Notebook/Magnetosome_Formation#title)
* [2014 Cornell](https://2014.igem.org/Team%3ACornell/notebook)

[![](https://static.igem.org/mediawiki/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:Hong Kong HKU/Engineering

HKU EVOLVE - Teams

[![](https://static.igem.org/mediawiki/2020/7/72/T--Hong_Kong_HKU--finfin.svg)](https://2020.igem.org/Team%3AHong_Kong_HKU)

Toggle navigation

* [Home](https://2020.igem.org/Team%3AHong_Kong_HKU)
* Team
  + [Team](https://2020.igem.org/Team%3AHong_Kong_HKU/Team)
  + [Attributions](https://2020.igem.org/Team%3AHong_Kong_HKU/Attributions)
* Project
  + [Background](https://2020.igem.org/Team%3AHong_Kong_HKU/Background)
  + [Description](https://2020.igem.org/Team%3AHong_Kong_HKU/Description)
  + [Results](https://2020.igem.org/Team%3AHong_Kong_HKU/results)
  + [Literature review](https://2020.igem.org/Team%3AHong_Kong_HKU/wetlabreview)
* Parts
  + [Part Collections](https://2020.igem.org/Team%3AHong_Kong_HKU/Part_Collection)
  + [Safety](https://2020.igem.org/Team%3AHong_Kong_HKU/safety)
* Modelling
  + [Dry Lab Overview](https://2020.igem.org/Team%3AHong_Kong_HKU/dryoverview)
  + [Simple Circuit](https://2020.igem.org/Team%3AHong_Kong_HKU/drylab)
  + [Release Assumptions](https://2020.igem.org/Team%3AHong_Kong_HKU/releasea)
  + [Kill Switch](https://2020.igem.org/Team%3AHong_Kong_HKU/killswitch)
  + [Program](https://2020.igem.org/Team%3AHong_Kong_HKU/program)
  + [Coding Details](https://2020.igem.org/Team%3AHong_Kong_HKU/learnmore)
  + [More Trials](https://2020.igem.org/Team%3AHong_Kong_HKU/moretrials)
  + [Dry Lab Review](https://2020.igem.org/Team%3AHong_Kong_HKU/dryreview)
* Human Practices
  + [Overview](https://2020.igem.org/Team%3AHong_Kong_HKU/HumanPractice)
  + [Integrated HP](https://2020.igem.org/Team%3AHong_Kong_HKU/meetings)
  + [Symposium](https://2020.igem.org/Team%3AHong_Kong_HKU/symposium)
  + [Public Engagement](https://2020.igem.org/Team%3AHong_Kong_HKU/education)
* Judging
  + [Medal Criteria](https://2020.igem.org/Team%3AHong_Kong_HKU/medalcriteria)
* Notebook
  + [Protocols](https://2020.igem.org/Team%3AHong_Kong_HKU/protocol)
  + [Logbook](https://2020.igem.org/Team%3AHong_Kong_HKU/Logbook)

# ENGINEERING ADVANCES

---

[

Your browser does not support the video tag.
](https://static.igem.org/mediawiki/2020/d/d0/T--Hong_Kong_HKU--modelintro.mp4)

Through our iGEM project, we have demonstrated the proof-of-concept of our novel Cre-loxp circuit through computational modelling and showed that our system can produce proteins in a defined ratio with high stability. We adopted the engineering cycle to perfect our system through the process as follows:

![](https://static.igem.org/mediawiki/2020/a/ac/T--Hong_Kong_HKU--engcycle.png)

Research: Biomanufacturing industry has a need to produce proteins at defined ratios. Research on microbial bioprocessing advances. [(See more...)](https://2020.igem.org/Team%3AHong_Kong_HKU/wetlabreview)

Imagine: Can we use a single microorganism that can express two or more phenotypes for different protein production?

Design: Develop the cre-loxp circuit design. [(See more...)](https://2020.igem.org/Team%3AHong_Kong_HKU/Description)

Build: Construct the computational model to simulate the cell growth kinetics and phenotype switching dynamics. [(See more...)](https://2020.igem.org/Team%3AHong_Kong_HKU/program)

Test: Run the model and obtain the results. [(See more...)](https://2020.igem.org/Team%3AHong_Kong_HKU/drylab)

Learn: Realised that our system can produce phenotypes at a constant ratio in the long run with very low variations. [(See more...)](https://2020.igem.org/Team%3AHong_Kong_HKU/drylab)

Improve: Release the assumptions in the computational model [(See more...)](https://2020.igem.org/Team%3AHong_Kong_HKU/releasea)

As we do not have full access to laboratory due to the COVID situation, we mainly put our focus on modelling and program development.

With limited time for wet lab, we were able to:- #### Design parts
- #### Amplification of parts
- #### 5 Site-directed mutagenesis on Cre recombinase
- #### Rounds of Lox-reporter construct and Cre construct assembly
- #### Co-transformation and pilot experiment
- #### In vitro Wild-Type Cre shuffling
- #### Multiple rounds of In vivo Wild-Type Cre shuffling with optimisation of parameters

**Learn more about our models and program by clicking on the cards below!**

![](https://static.igem.org/mediawiki/2020/f/f2/T--Hong_Kong_HKU--b1.png)

##### **Simple Circuit Model**

Our model finds:
(1) Stable phenotype over time with low coefficient of variations
(2) Phenotype population ratio can be controlled by the metabolic burden and the probabilities of phenotype switching of the two phenotypes

![](https://static.igem.org/mediawiki/2020/a/a4/T--Hong_Kong_HKU--b2.png)

##### **Programming**

We developed a database in which users can input their desired ratio and metabolic burden to generate the required probabilities of phenotype switching among two phenotypes that can be controlled in vitro

 ![](https://static.igem.org/mediawiki/2020/3/34/T--Hong_Kong_HKU--b3.png)

##### **Literature**

We believe that we can promote Synthetic Biology to the new generation of scientists, and students from all backgrounds. We therefore tutored 2 high school iGEM teams, and hosted an online course open to all undergraduates.

---

[![](https://static.igem.org/mediawiki/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:Hong Kong HKU/Experiments

# Experiments

Describe the research, experiments, and protocols you used in your iGEM project. These should be detailed enough for another team to repeat your experiments.

If you made Parts this year, please remember to put all information, characterization, and measurement data on the Part's Main Page on the [Registry](http://parts.igem.org/Main_Page).

### What should this page contain?

* Protocols
* Experiments

### Inspiration

* [2014 Colombia](https://2014.igem.org/Team%3AColombia/Protocols)
* [2014 Imperial](https://2014.igem.org/Team%3AImperial/Protocols)
* [2014 Caltech](https://2014.igem.org/Team%3ACaltech/Project/Experiments)

[![](https://static.igem.org/mediawiki/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:Hong Kong HKU/Results

HKU EVOLVE - RESULTS

[![HKU LOGO](https://static.igem.org/mediawiki/2020/d/d2/T--Hong_Kong_HKU--LOGO.png)](https://2020.igem.org/Team%3AHong_Kong_HKU)

Toggle navigation

* [Home](https://2020.igem.org/Team%3AHong_Kong_HKU)
* Team
  + [Team](https://2020.igem.org/Team%3AHong_Kong_HKU/Team)
  + [Attributions](https://2020.igem.org/Team%3AHong_Kong_HKU/Attributions)
* Project
  + [Background](https://2020.igem.org/Team%3AHong_Kong_HKU/Background)
  + [Description](https://2020.igem.org/Team%3AHong_Kong_HKU/Description)
  + [Results](https://2020.igem.org/Team%3AHong_Kong_HKU/results)
  + [Literature review](https://2020.igem.org/Team%3AHong_Kong_HKU/wetlabreview)
* Parts
  + [Part
    Collections](https://2020.igem.org/Team%3AHong_Kong_HKU/Part_Collection)
  + [Safety](https://2020.igem.org/Team%3AHong_Kong_HKU/safety)
* Modelling
  + [Dry
    Overview](https://2020.igem.org/Team%3AHong_Kong_HKU/dryoverview)
  + [Release
    Assumptions](https://2020.igem.org/Team%3AHong_Kong_HKU/releasea)
  + [Kill
    Switch](https://2020.igem.org/Team%3AHong_Kong_HKU/killswitch)
  + [Program](https://2020.igem.org/Team%3AHong_Kong_HKU/program)
  + [Coding
    Details](https://2020.igem.org/Team%3AHong_Kong_HKU/learnmore)
  + [Simple
    Circuit](https://2020.igem.org/Team%3AHong_Kong_HKU/drylab)
  + [More
    Trials](https://2020.igem.org/Team%3AHong_Kong_HKU/moretrials)
  + [Dry Lab
    Review](https://2020.igem.org/Team%3AHong_Kong_HKU/dryreview)
* Human Practices
  + [Overview](https://2020.igem.org/Team%3AHong_Kong_HKU/HumanPractice)
  + [Integrated
    Human](https://2020.igem.org/Team%3AHong_Kong_HKU/meetings)
  + [Symposium](https://2020.igem.org/Team%3AHong_Kong_HKU/symposium)
  + [Public
    Engagement](https://2020.igem.org/Team%3AHong_Kong_HKU/education)
* Judging
  + [Medal
    Criteria](https://2020.igem.org/Team%3AHong_Kong_HKU/medalcriteria)
* Notebook
  + [Protocols](https://2020.igem.org/Team%3AHong_Kong_HKU/protocol)
  + [Logbook](https://2020.igem.org/Team%3AHong_Kong_HKU/Logbook)
* Q&A

# RESULTS - WET LAB

![](https://static.igem.org/mediawiki/2020/6/62/T--Hong_Kong_HKU--wiki3445j.png)

### The Design

After we identified our research goal, which is to achieve a control over the ratiometric distribution
of two or more phenotypes of bacteria within a culture, we made use of the inversion property of Cre-lox
and avoided any deletion event in order to preserve all of the possible phenotypes in the gene pool.

We designed the lox-reporter constructs and cre mutant constructs with Snapgene and Geneious Prime.
Figure 1 illustrates the general structure of the lox-reporter construct. For easier observation of the
flipping events, we used fluorescent reporter genes such as green fluorescent protein (GFP) and red
fluorescent protein (RFP) for colorimetric analysis. To minimize the metabolic burden given by the
lox-reporter construct, we placed an inducible promoter upstream of the first lox sites.

![](https://static.igem.org/mediawiki/2020/6/67/T--Hong_Kong_HKU--1_reporter.png)

Figure 1: Design of lox-reporter construct. In the expression cassette, the orthogonal lox sites
sandwich two distinctive sets of reporter gene cassettes. GFP and RFP genes are in different
directions,
so only one gene will be expressed at a time. For irreversible recombination events, GFP (forward)
will
be expressed in the absence of Cre whereas RFP will instead be expressed after being flipped by Cre.

On the other hand, we also created a variety of cre mutants by site-specific mutagenesis, with reference
to Dr Eroshenko's paper.1In the pursuit of improving the specificity of the Cre-based recombination, we
used various Cre mutants in our project, instead of the wild types. The cre recombinase mutants are
designed to have one amino acid difference as compared to the wildtype. The change in amino acid
sequence may slightly alter the cre conformation, in particular the active sites. The cre mutants used
include Cre-R32V, Cre-R32M, 303GVSdup and Cre-Y324F.

With Pymol, the wild-type Cre recombinase and all the mutated versions (specific residues highlighted)
could be visualized. Figure 2 shows the wild-type Cre recombinases which form a dimer around a strand of
DNA. Figure 3 shows all the Cre mutants used in our experiment (R32M, R32V, Y324F and 303GVSdup).
According to the paper by Dr Eroshenko, three of the mutants (R32M, R32V and 303GVSdup) shows higher
specificity as the mutation destabilizes the linkage between the 2 dimers. As the authors suggest,
instead of disturbing the DNA-protein interaction, dimer interaction is a key to Cre recombinase
specificity. We did a meeting with Dr Eroshenko discussing about his paper and our project [Link Here](https://2020.igem.org/Team%3AHong_Kong_HKU/meetings)

---

![](https://static.igem.org/mediawiki/2020/4/40/T--Hong_Kong_HKU--Cre_WT_dimer_DNA.png)

Figure 2: 3D structure of Cre recombinase dimer interacting with a strand of DNA. Pymol was used to
visualize the crystal structure of specific protein with PDB files available in online databases.

![](https://static.igem.org/mediawiki/2020/c/c8/T--Hong_Kong_HKU--Cre333.png)

Figure 3: 3D structure of Cre recombinase mutants. A-B: Cre R32M mutant; C: 303GVSdup mutant
(mutated segment hightlighted in white circle); D-E: Cre R32V mutant; F: Y324F mutant (mutated
residues hightlighted in white circle). As the structure and residues change suggest, destabilizing
dimer interaction could enhance specificity.

In addition to the mutations created on Cre recombinases, variation is also added to the ribosome
binding site (RBS). A stronger RBS refers to a more efficient translation of the mRNA and therefore high
expression level of Cre. In this project, strong and weak RBS are incorporated to the Cre-mutant
construct for the determination of the metabolic burden and recombination efficiency.

![](https://static.igem.org/mediawiki/2020/a/a0/T--Hong_Kong_HKU--4_cre_construct.png)

Figure 4: Design of Cre-mutant constructs. The expression of Cre recombinase is controlled by the
level of arabinose. Mutation would also be introduced to the Cre gene for improving the specificity
of cre binding and in turns increasing recombination efficiency.

---

### The experiments

1. #### Amplification of parts

   The biobricks involved in this project are mainly from the iGEM distribution kit and IDT
   oligo. After we received the distribution kits, we first amplified the essential biobricks
   such as the lox sites and Cre-encoding sequences, as well as the backbones with different
   resistance and copy numbers. (state more details here first mb, like explain or show the
   Snapgene file for psb1c3)
2. #### Site-directed mutagenesis on Cre recombinase

   We used 4 sets of specific primers that would introduce desirable mutation to wild-type Cre
   during polymerase chain reaction (PCR). The mutations include point mutations (R32M, R32V
   and Y324F) and duplication (303GVSdup).

   Sanger sequencing in both forward and reverse directions showed that we successfully created
   four cre mutants which were then incorporated into the corresponding backbones containing
   ribosome binding sites (RBS) and arabinose-inducible promoter.The alignment result showed
   that all of the mutations had been successfully and accurately introduced to the Cre gene
   individually by site-directed mutagenesis.

   ![](https://static.igem.org/mediawiki/2020/5/59/T--Hong_Kong_HKU--5_seq.png)

   Figure 5: Multiple alignment between WT Cre and Cre mutants. A three-nucleotide change was
   introduced to the Cre gene which will encode for different amino acids at position 32 of the
   protein sequence.
3. #### Lox-reporter construct and Cre construct assembly

   Each biobrick contains a prefix (containing EcoRI and XbaI restriction sites) and suffix
   (containing SpeI and PstI restriction sites) and has been arranged in a standard psB1C3
   plasmid. We extracted the biobricks with appropriate restriction enzymes and assembled them
   through DNA ligation.

   We assembled the lox-reporter constructs and the cre mutant constructs by stepwise molecular
   cloning -- adding the ribosome binding site (RBS), the promoters and the reporter genes and
   the terminators, bit-by-bit into the psB1C3 backbone (Figure 1 and Figure 2). The
   recombinant plasmid was then transformed into the DH5a E. Coli strain and the selected
   colonies went through colony PCR and was amplified by bacterial culture. The transformants
   were verified by restriction enzyme digestion followed by agarose gel electrophoresis. Only
   when the restriction fragments match the expected size of the vector and that of the
   inserted gene could the cloning be said to be successful (figure 7). The presence of two
   fragments (insert & backbones) in the lane indicates the successful digestion of the
   construct. The respective size of the fragments further implies the presence of all
   essential components in the construct. All of the cre mutants were correctly cloned with all
   of the essential components. However, only loxP and lox2272 among all of the lox-reporter
   constructs showed the correct fragment sizes.

   ![](https://static.igem.org/mediawiki/2020/5/52/T--Hong_Kong_HKU--9_cloningflow.png)

   Figure 6: Schematic of the stepwise cloning

   ---

   ![](https://static.igem.org/mediawiki/2020/0/04/T--Hong_Kong_HKU--5_loxgel.png)
   ![](https://static.igem.org/mediawiki/2020/e/e6/T--Hong_Kong_HKU--6_cregel.png)

   Figure 7: Gel electrophoresis for digestion products of lox-reporter constructs (Left) and
   cre-mutant constructs (Right).
4. #### Switching backbones

   We created lox-reporter constructs and Cre-mutant constructs in psB1C3 backbone which is a
   high copy number plasmid containing chloramphenicol resistance. As double antibiotics
   resistance is required for co-transformation at the later stage for testing the ratiometric
   control of lox-cre recombination, we needed to switch the cre-mutant and lox-reporter to
   different backbones. We switched the lox-reporter construct to pSB1A3 (high copy number,
   ampicillin resistance). For Cre mutants, we selected plasmids with chloramphenicol
   resistance and assigned the copy number according to the strength of the RBS -- strong RBS
   with low copy number plasmid (i.e. psB4C5) and weak RBS with high copy number plasmid (i.e.
   psB1C3). This is to even out the final expression level between Cre-mutant constructs with
   strong RBS and those with weak RBS to avoid metabolic overload to the competent cells.
5. #### Co-transformation and first experiment

   For the validation of our lox-Cre system, we intend to first co-transform the lox-reporter
   and cre mutants into the competent cells for lox-cre recombination to occur, and then induce
   the Cre expression with different concentration of arabinose. The validation ends by
   re-transforming the lox-reporter to visualize the colour of the colonies. The ratio of Green
   colonies: Red colonies would imply the Cre-lox recombination efficiency. (However, due to
   the COVID-19 outbreak, the wetlab progress remains stagnant and real experiment could not be
   done properly.)
6. #### In vitro Wild-Type Cre shuffling

   ![](https://static.igem.org/mediawiki/2020/2/25/T--Hong_Kong_HKU--7_flip.png)

   Figure 8: Showing the gel of reporter construct with restriction digestion checking of (a)
   Deletion events (EcoRI, PstI E+P), and (b) Flipping Events (EcoRI, BsrGI E+B). Also
   displaying the copy number variation for the time to reach complete equilibrium with
   variation of substrate DNA amount.

   The Cre shuffling was conducted with 1 unit of Wild Type Cre in 37C for 30 minutes, with
   125ng of template as 1X substrate. With a wild type Cre, there is a miscellaneous band at
   ~750bp, which indicates the loxP sites interacting and recombining with a plasmid backbone
   sequence. If it is a deletion due to 2 loxP sites, the expected size would be 260bp with
   EcoI and PstI digestion, which is not present in the gel. It indicates the deletion is
   likely to be with an off-target site. This re-asserts the importance of the use of mutant
   versions of Cre in our system, as discussed in the meeting with Dr Eroshenko.

   With Image J, we could calculate the amount of flipped construct versus initial construct in
   1X substrate.

   | Band | Intensity | Length | Intensity/Length | Percentage of Inversion |
   | --- | --- | --- | --- | --- |
   | E+B Initial Backbone | 33727 | 3662 | 9.21 | 50.547% |
   | E+B Inverted Backbone | 19397 | 2476 | 7.83 | / |
    E+B Inverted Fragment | 19327 | 1758 | 10.99 | / |Table 1: Ratio of unflipped construct to flipped construct

   Therefore, we could see it reach equilibrium state within 30 minutes.

   Our reporter construct used in the pilot testing would be a high-copy number plasmid
   (100-300 copies) which directly extends the time to reach equilibrium, as illustrated by
   this experiment. With a normal 1 unit Cre with 0.125 ug construct with 2 loxP sites, the
   shuffling went to equilibrium state. However, the others with 10X or 100X copy number, there
   is no sign of flipped construct if equal DNA amount is loaded to gel, indicating much slower
   progress for all to equilibrium position of 50%.

   Hence, if we were to use the shuffling system with a single copy gene introduced with a
   lambda red system, it will only need a hundredth of the time needed to reach equilibrium.
   Therefore, it also provides good dry lab data to simulate rate and probability of flipping
   with a high-copy number plasmid model, after in vivo testing.
7. #### In vivo Wild-Type Cre shuffling

   ![](https://static.igem.org/mediawiki/2020/9/96/T--Hong_Kong_HKU--10_invivo.png)

   Figure 9: Showing the restriction digestion results of plasmids extracted after arabinose
   induction. EP indicates EcoRI + PstI digestion, and EB indicates EcoRI + BsrGI digestion. WT
   = Cre wild type. R32V, R32M, 303dup = Cre mutants with high specificity. Y324F = non-active
   Cre mutants.

   The reporter construct and the Cre constructs (high copy, weak RBS) was co-transformed into
   NEB stable E. Coli cell. The culture was grown in 0.2% arabinose with cells extracted at 30
   minutes, 2.5 hour, 4 hour, 16 hour, 24 hour and 48 hour, in presence of chloramphenicol and
   carbenicillin. Cells extracted were spin down, washed twice and incubated with medium with
   carbenicillin in LB, growing until the 24 hour mark. Plasmids were extracted with a
   mini-prep kit and analysed with restriction digestion.

   From the result, we could see mutant versions have less genome targeting activity in a same
   time-frame. From the left, we could see the deletion band (Red 1), the absence of the
   deletion band indicates the complete loss of the biobrick in the vector, since it represents
   the EcoRI + PstI digestion. It indicates the integration of the whole reporter plasmid into
   the bacterial genome. From the WT EB cut, we could also see the absence of the reporter
   plasmid in the Red 2. It shows the instability of maintaining lox construct in genome level
   or plasmid with wild type Cre, due to the cross-reactivity with genome off-target sites.

   Cre-mutants show a much lower extent of genome off-target effect. However, due to possible
   over-expression of Cre which leads to excision instead of inversion, we could not observe
   correct inversion patterns (Green box), but only excision patterns (Red 1) in EP cut. The
   non-cutting control shows correct banding of the initial position cut (Blue box).

   This shows we will need to adjust and lower the Cre expression level, since from the
   original Cre-mutant paper, the inversion could be maintained without detectable excision
   pattern [1]. Studies also show with too much Cre inside a system, deletion would occur
   instead of inversions by losing the directionality of lox sites, which leads to our
   observation[2]. Even with a weak RBS, the cre expression is too high, therefore the approach
   would be to decrease the arabinose concentration to further tune and reduce cre mutant
   expression.

   This also gives an initiative to investigate into other recombinase systems which does not
   involve deletion, including Hin, Gin, Fin orthogonal systems, which could use the same model
   and design to achieve the same goal.

---

### The future

There are few things that we want to accomplish in coming year:

1. Prove that our system works by completing the co-transformation and analyse the green: red colonies
   ratio.
2. Get into a more complex design of the lox-Cre recombinase.

   ![](https://static.igem.org/mediawiki/2020/7/71/T--Hong_Kong_HKU--8_complex.png)

   Figure 10: A more complex design of our lox-cre system.
3. crRNA cas13 designs to test and eliminate intermediate zones.

---

### References:

1. Eroshenko N, Church GM. Mutants of Cre recombinase with improved accuracy. Nature communications.
   2013 Sep 23;4(1):1-0.
2. Campo N, Daveran-Mingot ML, Leenhouts K, Ritzenthaler P, Le Bourgeois P. Cre-loxP recombination
   system for large genome rearrangements in Lactococcus lactis. Applied and Environmental
   Microbiology. 2002 May 1;68(5):2359-67.

##### HKU EVOLVE

We are HKU iGEM 2020 Team. We strive to establish ratiometric control over monocultures
of cells to achieve multicellular cooperation

##### Quick Links

* [Home](https://2020.igem.org/Team%3AHong_Kong_HKU)
* [Team](https://2020.igem.org/Team%3AHong_Kong_HKU/Team)
* [Project](https://2020.igem.org/Team%3AHong_Kong_HKU/Background)
* [Parts](https://2020.igem.org/Team%3AHong_Kong_HKU/Part_Collection)
* [Modelling](https://2020.igem.org/Team%3AHong_Kong_HKU/dryoverview)
* [Human
  Practices](https://2020.igem.org/Team%3AHong_Kong_HKU/HumanPractice)
* [Judging](https://2020.igem.org/Team%3AHong_Kong_HKU/medalcriteria)

##### Our Sponsors

* ![idt](https://static.igem.org/mediawiki/2020/b/b9/T--Hong_Kong_HKU--idt.png)
* ![bench](https://static.igem.org/mediawiki/2020/f/ff/T--Hong_Kong_HKU--bench.png)
* ![neb](https://static.igem.org/mediawiki/2020/e/ea/T--HK_SSC--NEB.png)
* ![twist](https://static.igem.org/mediawiki/2020/6/62/T--Hong_Kong_HKU--twistf.png)

##### Contact Us

* ![email](https://static.igem.org/mediawiki/2020/e/e3/T--Hong_Kong_HKU--mail.png)
* [![Facebook](https://static.igem.org/mediawiki/2020/0/0e/T--Hong_Kong_HKU--Facebookf.png)](https://m.facebook.com/hkuigem2020/ "Facebook")
* [![instagram](https://static.igem.org/mediawiki/2020/3/3a/T--Hong_Kong_HKU--igf.png)](https://www.instagram.com/hkuevolve/ "instagram")

HKU iGEM Team 2020