A New Method for the Modification of Fibroin Heavy Chain Protein in the Transgenic Silkworm

Kojima, Katsura; Kuwana, Yoshihiko; Sezutsu, Hideki; Kobayashi, Isao; Uchino, Keiro; Tamura, Toshiki; Tamada, Yasushi

doi:10.1271/bbb.70353

Cited by 60 publications

(74 citation statements)

References 29 publications

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“…Silkworms can be efficiently transformed with piggyBac vectors (25,26), and piggyBac vectors can be engineered to target foreign gene expression in a silk gland-specific fashion (27)(28)(29) and to direct the incor- poration of foreign proteins into composite silk fibers (27)(28)(29)(30)32). The use of piggyBac vectors with these features has yielded transgenic silkworm lines that produced composite fibers containing a variety of different recombinant proteins, including procollagen (28), GFP (27,29), and feline IFN (30). In addition, while our work was in progress, others isolated transgenic silkworms that can produce spider silk proteins (35,36).…”

Section: Discussionmentioning

confidence: 99%

“…However, we also recognized that this protein would be expressed in concert and would have to interact with the endogenous silkworm fhc, fibroin light chain, and fibrohexamerin proteins to be incorporated into silk fibers. Thus, we embedded the A2S8 14 spider silk sequence within N-and C-terminal fhc domains to direct the assembly process, as originally described by Kojima and coworkers (29). Together with the ability of the fhc promoter to drive their expression in spatial and temporal proximity to the endogenous silkworm silk proteins, we believe that these design features accounted for the ability of the chimeric silkworm/spider silk proteins to participate in the assembly of composite silk fibers and contribute significantly to their mechanical properties.…”

Section: Discussionmentioning

confidence: 99%

“…Although silkworms are not recognized as a major recombinant protein production platform (21)(22)(23), they might be the perfect host for spider silk fiber production because transgenic silkworms can be efficiently produced using piggyBac vectors (24)(25)(26), recombinant protein production can be targeted to the silk gland with tissuespecific promoters (27)(28)(29)(30)(31)(32), and the silk gland is naturally equipped to assemble silk proteins into fibers (33). One caveat is that silkworms produce endogenous silk proteins.…”

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confidence: 99%

“…1). These included the Bombyx mori fibroin heavy chain (fhc) promoter to target expression of the foreign spider silk protein to the posterior silk gland (29,30) and an fhc enhancer to increase expression levels to facilitate assembly of the foreign silk protein into fibers (31). Our piggyBac vectors also encoded A2S8 14 ( Fig.…”

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confidence: 99%

“…1 B and C). This chimeric silkworm/spider silk design was used previously to direct foreign proteins into nascent, endogenous silk fibers in the B. mori silk gland to produce composite silk fibers (29,30). One of the piggyBac vectors constructed in this study encoded the chimeric silkworm/spider silk protein alone (Fig.…”

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Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties

Teulé

Miao

Sohn

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

241

158

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The development of a spider silk-manufacturing process is of great interest. However, there are serious problems with natural manufacturing through spider farming, and standard recombinant protein production platforms have provided limited progress due to their inability to assemble spider silk proteins into fibers. Thus, we used piggyBac vectors to create transgenic silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk proteins integrated in an extremely stable manner. Furthermore, these composite fibers were, on average, tougher than the parental silkworm silk fibers and as tough as native dragline spider silk fibers. These results demonstrate that silkworms can be engineered to manufacture composite silk fibers containing stably integrated spider silk protein sequences, which significantly improve the overall mechanical properties of the parental silkworm silk fibers.biomaterials | biotechnology | transgenic animals

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties

Teulé

Miao

Sohn

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

241

158

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Colored Fluorescent Silk Made by Transgenic Silkworms

Iizuka

Sezutsu

Tatematsu

et al. 2013

Adv Funct Materials

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Silk is a protein fiber used to weave fabrics and as a biomaterial in medical applications. Recently, genetically modified silks have been produced from transgenic silkworms. In the present study, transgenic silkworms for the mass production of three colors of fluorescent silks, (green, red, and orange) are generated using a vector originating from the fibroin H chain gene and a classical breeding method. The suitability of the recombinant silks for making fabrics is investigated by harvesting large amounts of the cocoons, obtained from rearing over 20 thousand silkworms. The application of low temperature and a weakly alkaline solution for cooking and reeling enables the production of silk fiber without loss of color. The maximum strain tolerated and Young's modulus of the fluorescent silks are similar to those of ordinary silk, although the maximum stress value of the recombinant silk is slightly lower than that of the control. Fabrics with fluorescent color are demonstrated using the recombinant silk, with the color persisting for over two years. The results indicate that large amounts of genetically modified silk can be made by transgenic silkworms, and the silk is applicable as functional silk fiber for making fabrics and for use in medical applications.

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Biomedical Applications of Recombinant Silk‐Based Materials

Aigner

DeSimone

Scheibel³

2018

Advanced Materials

243

211

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Silk is mostly known as a luxurious textile, which originates from silkworms first cultivated in China. A deeper look into the variety of silk reveals that it can be used for much more, in nature and by humanity. For medical purposes, natural silks were recognized early as a potential biomaterial for surgical threads or wound dressings; however, as biomedical engineering advances, the demand for high-performance, naturally derived biomaterials becomes more pressing and stringent. A common problem of natural materials is their large batch-to-batch variation, the quantity available, their potentially high immunogenicity, and their fast biodegradation. Some of these common problems also apply to silk; therefore, recombinant approaches for producing silk proteins have been developed. There are several research groups which study and utilize various recombinantly produced silk proteins, and many of these have also investigated their products for biomedical applications. This review gives a critical overview over of the results for applications of recombinant silk proteins in biomedical engineering.

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A New Method for the Modification of Fibroin Heavy Chain Protein in the Transgenic Silkworm

Cited by 60 publications

References 29 publications

Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties

Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties

Colored Fluorescent Silk Made by Transgenic Silkworms

Biomedical Applications of Recombinant Silk‐Based Materials

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