Project

Introduction

CAFA-Beijing is a team composed entirely of faculty and students from art-related disciplines. So why are we participating in iGEM? Because synthetic biology may well be one of the most effective pathways through which we can translate cutting-edge art into tangible benefits for all of humanity. While we are passionate about bio-art, we have deliberately chosen to pursue a therapy-oriented project. This bold decision reflects our aspiration to engage in a grand interdisciplinary dialogue — one that moves beyond using synthetic biology merely as a showcase for artistic expression, but rather integrates an artistic mindset into technological development that can more rapidly impact people's daily lives. The term 'cosmetic medicine' inherently embodies both 'medicine' and 'aesthetics' — which is precisely why CAFA-Beijing has initiated the ChronoKolla project. We may not achieve perfection at this stage, but we are committed to taking this first step with conviction and courage.

Project Context

Three thousand years ago in Egypt, queens used black seed oil to protect their hair and relied on its antioxidant and antibacterial properties to preserve youth.
Two thousand years ago in China, the Shennong Herbal Classic documented more than 160 herbs with beauty-enhancing effects. For example, Bombyx Batryticatus was believed to "darken the hair and enhance appearance," while Radix Angelicae Dahuricae and Semen Benincasae were said to "nourish complexion," highlighting the importance of both oral remedies and topical applications in improving skin health.

However, the ultimate challenge in skincare—simultaneously addressing hyperpigmentation and aging—remains a luxury. Central to this is collagen, a vital structural protein in the human body. Collagen is abundant in skin, bones, muscles, connective tissue, keratin, and even organs, making up about 30% of total human protein. It plays essential roles in maintaining skin elasticity, protecting organs, and supporting strength and flexibility.

Collagen peptides are smaller fragments of collagen. Supplementation—whether taken orally or applied externally—provides short peptide chains and amino acids that the body can repurpose to synthesize necessary proteins. For this reason, collagen peptides are widely incorporated into health and skincare products. However, because natural human- and animal-derived collagen peptides are limited, most commercial products rely on artificially recombinant collagen peptides. These recombinant versions often suffer from limited efficacy, high cost, and underwhelming results. In practice, their benefits are maximized mainly in beauty salon treatments rather than in everyday use.

Collagen skincare today generally falls into three approaches: topical application, oral supplementation, and beauty salon procedures. Faced with limited options, some people spend equal amounts of time and money—but at greater risk—to pursue plastic surgery, which seems more effective in the short term.

Ultimately, modern medical beauty has become a "triple luxury" of time, money, and risk. In the process, many seem to forget that skincare is a fundamental human need.

In one sentence, the mission of our project is simple: to give everyone the right to accessible, effective skincare.

Project Description

1. Engineering Bacteria
   To address the limited efficacy of existing recombinant collagen peptides, we designed a novel fusion peptide through computer-aided methods. This peptide analog competes with the melanocyte-stimulating hormone receptor (α-MSH), thereby blocking melanin synthesis and reducing pigmentation. Additionally, fragments mimicking growth factors were incorporated into the fusion sequence to stimulate skin cells, delay aging, and promote collagen production.
   
   
   

   

   
   To enhance cellular uptake, a cell-penetrating peptide was added to the C-terminus of the fusion peptide, enabling efficient entry into skin cells and ensuring biological activity. We plan to express these fusion peptides using E. coli BL21.
   
   
   Finally, to overcome the high production costs associated with current collagen peptides, we integrated multiple functions—melanin inhibition, anti-aging activity, and cell penetration—into a single peptide sequence. By encoding this sequence into one plasmid and introducing it into engineered bacteria, we avoid the need to cultivate multiple strains separately. This strategy increases production efficiency, enhances peptide yield, and significantly reduces manufacturing costs.
   
   
2. Hardware
   To ensure effective delivery, we designed a microneedle-based substrate system capable of transporting active compounds directly into the dermis. This method improves bioavailability and maximizes the therapeutic effects of the engineered peptides.
   
   
   

   

Project Implementation

We completed the entire process from gene design to functional validation through a systematic experimental workflow. The project team worked collaboratively with clear task division, ensuring that each stage achieved its expected objectives and laid a solid foundation for subsequent applications.

The engineered bacteria primarily express three classes of peptides.

Engineering bacteria

First, we amplified the designed plasmid using Escherichia coli DH5α, and then introduced it into endotoxin-free Escherichia coli BL21 for expression.

1、a-MSH antagonist peptide

Gene1 encodes an a-MSH antagonist peptide, which blocks the binding of a-MSH to MC1R, thereby inhibiting melanin synthesis.

2、TGF-β mimetic peptide

Gene2 fragment encodes a peptide that mimics TGF-β,activating the Smad2/3 signaling pathway and stimulating fibroblasts to secrete collagen.

3、TAT cell-penetrating pepitide

In the TGF-β mimetic peptide, we have also incorporated gene3，a TAT transduction peptide, which serves as a delivery module to facilitate the transport of other peptides across intercellular spaces.

Feasibility

Technology

From a technological perspective, this project has a solid experimental foundation and a clear R&D method. To address the problem of the single efficacy of the existing recombinant collagen peptide, and to inhibit the production of melanin in skin cells. We utilized computer-aided methods to design a peptide analogue that competes with the melanocyte-stimulating hormone receptor (α-MSH), and this analogue can block the synthesis of melanin. To delay skin aging and promote collagen synthesis, we added fragments of simulated growth factors into the sequence of the fusion peptide. These sequence fragments can stimulate skin cells to produce collagen.

We added a segment of cell-penetrating peptide at the end of the fusion peptide to facilitate the entry of these compounds into skin cells, achieving their effective action. We plan to use Escherichia coli N1917 (ECN1917) to produce these fusion peptides.

By using genetic engineering techniques, a multifunctional fusion peptide with the functions of melanin inhibition, collagen promotion and cell penetration was designed. And the single-plasmid dual-fragment expression technology was adopted to increase the yield. This precisely addresses the industry pain points of existing recombinant collagen peptide having a single efficacy and high cost.

To sum up, the experimental process is carried out in three stages (plasmid construction → expression verification → functional testing), covering key points such as cytotoxicity, melanin inhibition rate, and collagen synthesis efficiency. The microneedle delivery system is used to enhance the skin absorption efficiency. The technical solution is mature and the risks are controllable.

Market

From a market perspective, the development of this collagen technology has significantly reduced production costs and facilitated the rapid expansion of the market. The medical aesthetic collagen peptide market is large, but products that combine "Anti-pigmentation ＆ Anti-aging" dual effects are scarce. The differentiated competitive advantage is remarkable and indicative. The growth of this sub-market is conducive to investment and development, as well as meeting the needs of consumers and industries. We also designed a delivery system based on micrometer-sized micro-needles, to achieve effective and simple administration of these compounds and the new product has significantly enhanced their portability.

Society

From a societal benefit perspective, the project integrates science communication with art experience innovatively. To eliminate public misunderstandings about collagen peptides, we aimed to "visualize" and "make tangible" the scientific principles in cross-border art venues such as the 798 Art District in Beijing through the popular science lecture "The Counter-Clockwise Code of the Human Body" and the multi-sensory interactive device that simulates the expression of genes and protein synthesis through light, sound and visual interaction. This artistic approach not only enhances public trust in biotechnology, but also meets the demands of the public for functional skin care products. As a result, it is easy to obtain policy and economic support.

Reference

Description

Lee M, Kim E, Ahn H, Son S, Lee H. Oral intake of collagen peptide NS improves hydration, elasticity, desquamation, and wrinkling in human skin: a randomized, double-blinded, placebo-controlled study. Food Funct. 2023 Apr 3;14(7):3196-3207. doi: 10.1039/d2fo02958h. PMID: 36916504.

Kviatkovsky SA, Hickner RC, Cabre HE, Small SD, Ormsbee MJ. Collagen peptides supplementation improves function, pain, and physical and mental outcomes in active adults. J Int Soc Sports Nutr. 2023 Dec;20(1):2243252. doi: 10.1080/15502783.2023.2243252. PMID: 37551682; PMCID: PMC10411303.

Avila Rodríguez MI, Rodríguez Barroso LG, Sánchez ML. Collagen: A review on its sources and potential cosmetic applications. J Cosmet Dermatol. 2018 Feb;17(1):20-26. doi: 10.1111/jocd.12450. Epub 2017 Nov 16. PMID: 29144022.

Zaelzer C. The Value in Science-Art Partnerships for Science Education and Science Communication. eNeuro. 2020 Jul 17;7(4):ENEURO.0238-20.2020. doi: 10.1523/ENEURO.0238-20.2020. PMID: 32616625; PMCID: PMC7369315.

Campos LD, Pereira ATSA, Cazarin CBB. The collagen market and knowledge, attitudes, and practices of Brazilian consumers regarding collagen ingestion. Food Res Int. 2023 Aug;170:112951. doi: 10.1016/j.foodres.2023.112951. Epub 2023 May 18. PMID: 37316004.

Design

Zhao L, Yin G, Zhang Y, Duan C, Wang Y, Kang Z. A comparative study on the genomes, transcriptomes, and metabolic properties of Escherichia coli strains Nissle 1917, BL21(DE3), and MG1655. Eng Microbiol. 2022 Feb 21;2(1):100012. doi: 10.1016/j.engmic.2022.100012. PMID: 39628614; PMCID: PMC11610980.

Szabó I, Biri-Kovács B, Vári B, Ranđelović I, Vári-Mező D, Juhász É, Halmos G, Bősze S, Tóvári J, Mező G. Targeting the Melanocortin 1 Receptor in Melanoma: Biological Activity of α-MSH-Peptide Conjugates. Int J Mol Sci. 2024 Jan 16;25(2):1095. doi: 10.3390/ijms25021095. PMID: 38256168; PMCID: PMC10816934.

Horinouchi CDS, Oostendorp C, Schade D, van Kuppevelt TH, Daamen WF. Growth factor mimetics for skin regeneration: In vitro profiling of primary human fibroblasts and keratinocytes. Arch Pharm (Weinheim). 2021 Aug;354(8):e2100082. doi: 10.1002/ardp.202100082. Epub 2021 May 8. PMID: 33963608.

Velimirovic M, Zanetti LC, Shen MW, Fife JD, Lin L, Cha M, Akinci E, Barnum D, Yu T, Sherwood RI. Peptide fusion improves prime editing efficiency. Nat Commun. 2022 Jun 18;13(1):3512. doi: 10.1038/s41467-022-31270-y. PMID: 35717416; PMCID: PMC9206660.

Kirkby M, Hutton ARJ, Donnelly RF. Microneedle Mediated Transdermal Delivery of Protein, Peptide and Antibody Based Therapeutics: Current Status and Future Considerations. Pharm Res. 2020 Jun 2;37(6):117. doi: 10.1007/s11095-020-02844-6. PMID: 32488611; PMCID: PMC7266419.

Nguyen NH, Nguyen TT, Bui VKH, Nguyen NTT, Van Vo G. Recent advances in microneedles for enhanced functional angiogenesis and vascular drug delivery. Ther Deliv. 2025 Apr;16(4):393-406. doi: 10.1080/20415990.2025.2468148. Epub 2025 Feb 25. PMID: 39997030; PMCID: PMC11970790.

Prausnitz MR. Engineering Microneedle Patches for Vaccination and Drug Delivery to Skin. Annu Rev Chem Biomol Eng. 2017 Jun 7;8:177-200. doi: 10.1146/annurev-chembioeng-060816-101514. Epub 2017 Mar 24. PMID: 28375775.

Dall'Olmo L, Papa N, Surdo NC, Marigo I, Mocellin S. Alpha-melanocyte stimulating hormone (α-MSH): biology, clinical relevance and implication in melanoma. J Transl Med. 2023 Aug 22;21(1):562. doi: 10.1186/s12967-023-04405-y. PMID: 37608347; PMCID: PMC10463388.

Rizzo MG, Palermo N, D'Amora U, Oddo S, Guglielmino SPP, Conoci S, Szychlinska MA, Calabrese G. Multipotential Role of Growth Factor Mimetic Peptides for Osteochondral Tissue Engineering. Int J Mol Sci. 2022 Jul 2;23(13):7388. doi: 10.3390/ijms23137388. PMID: 35806393; PMCID: PMC9266819.

Herraiz C, Martínez-Vicente I, Maresca V. The α-melanocyte-stimulating hormone/melanocortin-1 receptor interaction: A driver of pleiotropic effects beyond pigmentation. Pigment Cell Melanoma Res. 2021 Jul;34(4):748-761. doi: 10.1111/pcmr.12980. Epub 2021 May 2. PMID: 33884776.

Zorko M, Langel Ü. Cell-Penetrating Peptides. Methods Mol Biol. 2022;2383:3-32. doi: 10.1007/978-1-0716-1752-6_1. PMID: 34766279.

Chablani L, Singh V. Cell-Penetrating Peptides as Passive Permeation Enhancers for Transdermal Drug Delivery. AAPS PharmSciTech. 2022 Sep 26;23(7):266. doi: 10.1208/s12249-022-02424-4. PMID: 36163537.