title: Genome editing of BmFib-H gene provides an empty Bombyx mori silk gland for a highly efficient bioreactor.
abstract: Evolution has produced some remarkable creatures, of which silk gland is a fascinating organ that exists in a variety of insects and almost half of the 34,000 spider species. The impressive ability to secrete huge amount of pure silk protein, and to store proteins at an extremely high concentration (up to 25%) make the silk gland of Bombyx mori hold great promise to be a cost-effective platform for production of recombinant proteins. However, the extremely low production yields of the numerous reported expression systems greatly hindered the exploration and application of silk gland bioreactors. Using customized zinc finger nucleases (ZFN), we successfully performed genome editing of Bmfib-H gene, which encodes the largest and most abundant silk protein, in B. mori with efficiency higher than any previously reported. The resulted Bmfib-H knocked-out B. mori showed a smaller and empty silk gland, abnormally developed posterior silk gland cells, an extremely thin cocoon that contain only sericin proteins, and a slightly heavier pupae. We also showed that removal of endogenous Bmfib-H protein could significantly increase the expression level of exogenous protein. Furthermore, we demonstrated that the bioreactor is suitable for large scale production of protein-based materials.
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
authors.forename: Sanyuan
authors.initials: S
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
authors.forename: Run
authors.initials: R
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
authors.forename: Xiaogang
authors.initials: X
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
authors.forename: Jiasong
authors.initials: J
authors.lastname: Chang
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Lu
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
authors.forename: Jianduo
authors.initials: J
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
authors.forename: Qingyou
authors.initials: Q
authors.lastname: Xia
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D055786:Gene Knockout Techniques; D018390:Gene Targeting; D017344:Genes, Insect; D017930:Genes, Reporter; D049750:Genome, Insect; D023281:Genomics; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics; D016415:Sequence Alignment; D047011:Silk / Q000096:biosynthesis
chemical_list: D011993:Recombinant Fusion Proteins; D047011:Silk