Genetic hybridization of highly active exogenous functional proteins into silk-based materials using “light-clothing” strategy

Long, Dan; Cheng, Xian; Hao, Zhanzhang; Sun, Jianfeng; Diane, Umuhoza; Liu, Yanping; Chen, Lili; Zhang, Xiang; Yang, Fang; Zhao, Aichun

doi:10.1016/j.matt.2021.03.020

Cited by 16 publications

(11 citation statements)

References 50 publications

(89 reference statements)

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“…Figure 2A illustrates the preparation process of the SF microneedle under-layer. Since SF microneedles are used in the moist oral environment, an essential annealing step during preparation is to partially transform the SF molecules from a random coil to stable β-sheet conformation to render the SF water-insoluble ( Long D. et al, 2021 ; Long et al, 2021b ; Cheng et al, 2021 ). Three kinds of SF microneedles were constructed with low, medium and high β-sheet contents of 18.07% (low-β group), 24.86% (medium-β group) and 34.65% (high-β group), respectively ( Figures 2B, C ).…”

Section: Resultsmentioning

confidence: 99%

Drug-loaded mucoadhesive microneedle patch for the treatment of oral submucous fibrosis

Cheng,

Yang,

Liao

et al. 2023

Front. Bioeng. Biotechnol.

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Oral submucous fibrosis is a chronic, inflammatory and potentially malignant oral disease. Local delivery of triamcinolone to lesion site is a commonly used therapy. The existing methods for local drug delivery include topical administration and submucosal injection. However, in the wet and dynamic oral microenvironment, these methods have drawbacks such as limited drug delivery efficiency and injection pain. Therefore, it is urgently needed to develop an alternative local drug delivery system with high efficiency and painlessness. Inspired by the structure of band-aid, this study proposed a novel double-layered mucoadhesive microneedle patch for transmucosal drug delivery. The patch consisted of a mucoadhesive silk fibroin/tannic acid top-layer and a silk fibroin microneedle under-layer. When applying the annealing condition for the medium content of β-sheets of silk fibroin, the microneedles in under-layer displayed both superior morphology and mechanical property. The mechanical strength of per needle (0.071N) was sufficient to penetrate the oral mucosa. Sequentially, the gelation efficiency of silk fibroin and tannic acid in top-layer was maximized as the weight ratio of tannic acid to silk fibroin reached 5:1. Moreover, in vitro results demonstrated the double-layered patch possessed undetectable cytotoxicity. The sustained release of triamcinolone was observed from the double-layered patch for at least 7 days. Furthermore, compared with other commercial buccal patches, the double-layered patch exhibited an enhanced wet adhesion strength of 37.74 kPa. In addition, ex vivo mucosal tissue penetration experiment confirmed that the double-layered patch could reach the lamina propria, ensuring effective drug delivery to the lesion site of oral submucous fibrosis. These results illustrate the promising potential of the drug-loaded mucoadhesive microneedle patch for the treatment of oral submucous fibrosis.

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Section: Resultsmentioning

confidence: 99%

Drug-loaded mucoadhesive microneedle patch for the treatment of oral submucous fibrosis

Cheng,

Yang,

Liao

et al. 2023

Front. Bioeng. Biotechnol.

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“…2A). Therefore, we can introduce other tissuespecific promoter(s) (TSps), such as promoters specicf for silk-gland (Hao et al, 2021;Long et al, 2021), midgut (Jiang et al, 2013), hemocyte (Tsubota et al, 2014;Zhang et al, 2018), and fat body (Deng et al, 2013), to drive GAL4 gene expression cassette flanked by loxP/lox2272 sites that can be integrated into the genomes of activator strains via Cre-RMCE (Fig. 8A), and other genes of interest (GOIs) flanked by the attB/attB sites that can be integrated into the genomes of activator or effector strains via phiC31-RMCE (Fig.…”

Section: Discussionmentioning

confidence: 99%

An efficient and safe strategy for germ cell‐specific automatic excision of foreign DNA in F₁ hybrid transgenic silkworms

et al. 2023

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The safety of transgenic technology is a major obstacle in the popularization and use of transgenic silkworms and their products. In sericulture, only the first filial generation (F1) hybrid eggs produced by cross‐breeding Japanese and Chinese original strains are usually used for the large‐scale breeding of silkworms, but this may result in uncontrolled transgene dispersal during the popularization and application of the F1 hybrid transgenic eggs. To address this issue, we developed a safe and efficient strategy using the GAL4/Upstream activating sequence (UAS) system, the FLP/flippase recognition target (FRT) system, and the gonad‐specific expression gene promoters (RSHP1p and Nanosp) for the germ cell‐specific automatic excision of foreign DNA in the F1 hybrid transgenic silkworms. We established 2 types of activator strains, R1p::GAL4‐Gr and Nsp::GAL4‐Gr, containing the testis‐specific GAL4 gene expression cassettes driven by RSHP1p or Nanosp, respectively, and 1 type of effector strain, UAS::FLP‐Rg, containing the UAS‐linked FLP gene expression cassette. The FLP recombinase‐mediated sperm‐specific complete excision of FRT‐flanked target DNA in the F1 double‐transgenic silkworms resulting from the hybridization of R1p::GAL4‐Gr and UAS::FLP‐Rg was 100%, whereas the complete excision efficiency resulting from the hybridization of Nsp::GAL4‐Gr and UAS::FLP‐Rg ranged from 13.73% to 80.3%. Additionally, we identified a gene, sw11114, that is expressed in both testis and ovary of Bombyx mori, and can be used to establish novel gonad‐specific expression systems in transgenic silkworms. This strategy has the potential to fundamentally solve the safety issue in the production of F1 transgenic silkworm eggs and provides an important reference for the safety of transgenic technology in other insect species.

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“…CRISPR/Cas9-messenger RNP mixtures with volumes of approximately 1 nl were injected into the middle of the eggs, and the wound was sealed with glue. All the injected embryos were allowed to develop in a climate chamber at 25°C and 80% humidity ( Long et al, 2021 ).…”

Section: Methodsmentioning

confidence: 99%

The BTB-ZF gene Bm-mamo regulates pigmentation in silkworm caterpillars

Wu,

Tong,

Peng

et al. 2024

eLife

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Color pattern of insects is one of the most dazzling adaptive evolutionary phenotypes. However, the molecular regulation of this color pattern is not clear. In this paper, we found a transcription factor, Bm-mamo, is responsible for bd (black dilute) allele mutants in silkworm. It belongs to BTB zinc finger family, and is ortholog to mamo of Drosophila melanogaster, which It is found that this gene has conservative function in gamete production, and evolved a pleiotropic function in regulation of color patterns in caterpillar. We found that the Bm-mamo can comprehensively regulate the expression of related pigment synthesis and cuticular protein genes to form color patterns. This suggests that the deposition of pigment particles in caterpillars’ epidermis requires not only the spatiotemporal expression of pigment synthesis genes, but also the correct expression of related cuticular protein genes. This study provides a new data for the setting of color patterns.

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Genetic hybridization of highly active exogenous functional proteins into silk-based materials using “light-clothing” strategy

Cited by 16 publications

References 50 publications

Drug-loaded mucoadhesive microneedle patch for the treatment of oral submucous fibrosis

Drug-loaded mucoadhesive microneedle patch for the treatment of oral submucous fibrosis

An efficient and safe strategy for germ cell‐specific automatic excision of foreign DNA in F₁ hybrid transgenic silkworms

The BTB-ZF gene Bm-mamo regulates pigmentation in silkworm caterpillars

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Genetic hybridization of highly active exogenous functional proteins into silk-based materials using “light-clothing” strategy

Cited by 16 publications

References 50 publications

Drug-loaded mucoadhesive microneedle patch for the treatment of oral submucous fibrosis

Drug-loaded mucoadhesive microneedle patch for the treatment of oral submucous fibrosis

An efficient and safe strategy for germ cell‐specific automatic excision of foreign DNA in F1 hybrid transgenic silkworms

The BTB-ZF gene Bm-mamo regulates pigmentation in silkworm caterpillars

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An efficient and safe strategy for germ cell‐specific automatic excision of foreign DNA in F₁ hybrid transgenic silkworms