Transgenic silkworms expressing spider silk

Patent metadata

Field	Value
Publication number	US-11089767-B2
Application number	US-201815942185-A
Country	US
Kind code	B2
Filing date	Mar 30, 2018
Priority date	Mar 30, 2017
Publication date	Aug 17, 2021
Grant date	Aug 17, 2021

Abstract

Transgenic silkworms stably expressing synthetic spider silk genes or composite silkworm/spider silk genes are disclosed. The exogenous spider silk genes are stably intergrated into a defined site of the fibroin heavy chain intron or a fibroin light chain intron of silkworms. Synthetic spider silk proteins and composite spider silk-silkworm genes and proteins are provided. The expression of exogenous spider silk genes is driven by the endogenous fibroin heavy chain promoter, improving the genetic stability of transgenic silkworms. The composite silkworm/spider silk fibers exhibit exceptional mechanical performance, compared to normal silkworm silk fibers and other transgenic silkworm fibers.

First claim

What is claimed is:

1. A transgenic silkworm comprising a stably-integrated exogenous synthetic spider silk gene operably linked to an endogenous silkworm promoter, wherein the exogenous spider silk gene is greater than three kilobases (3 kb) in length, is stably integrated in a sixth intron of a fibroin light chain gene, FibL, wherein the endogenous silkworm promoter is a silkworm-specific U6 promoter and the transgenic silkworm stably expresses the spider silk gene.

2. The transgenic silkworm of claim 1 , wherein the exogenous synthetic spider silk gene is operably linked to an endogenous silkworm promoter in the silkworm gland.

3. The transgenic silkworm of claim 1 , wherein the exogenous spider silk gene is greater than four kilobases (4 kb) in length.

4. The transgenic silkworm of claim 1 , wherein the exogenous spider silk gene is greater than five kilobases (5 kb) in length.

5. The transgenic silkworm of claim 1 , wherein the exogenous spider silk gene is ten kilobases (10 kb) in length.

6. The transgenic silkworm of claim 1 , wherein the exogenous spider silk gene comprises SEQ ID NO. 25 or SEQ ID NO:26 or a fragment of either thereof.

7. The transgenic silkworm of claim 1 , wherein the transgenic silkworm stably expresses the spider silk gene.

8. The transgenic silkworm of claim 1 , wherein the exogenous synthetic spider silk gene is stably integrated in an intron of a fibroin heavy chain gene, FibH.

9. The transgenic silkworm of claim 1 , wherein the exogenous spider silk gene comprises the MaSp1 gene, MaSp2 gene, Ac1 gene, Flag gene, Piri gene or the MiSp1 gene of Nephila clavipes.

10. The transgenic silkworm of claim 1 , wherein the silkworm is Bombyx sp.

11. The transgenic silkworm of claim 10 , wherein the silkworm is Bombyx mori.

12. A progeny silkworm of the transgenic silkworm claim 1 , wherein the exogenous synthetic spider silk gene is stably integrated.

13. A transgenic Bombyx silkworm, the silkworm comprising:
a stably-integrated MaSp1 gene or the MiSp1 gene of Nephila clavipes synthetic spider silk gene in the intron of the fibroin heavy chain gene FibH and operably linked to the silkworm-specific U6 promoter,
wherein the exogenous spider silk gene is greater than three kilobases (3 kb) in length,
wherein the transgenic silkworm stably expresses the spider silk gene.

14. The transgenic silkworm of claim 13 , wherein the exogenous spider silk gene comprises SEQ ID NO. 25 or SEQ ID NO:26, or a fragment of either thereof.

15. A transgenic silkworm comprising a stably-integrated exogenous synthetic spider silk gene operably linked to an endogenous silkworm promoter, wherein the exogenous spider silk gene is greater than three kilobases (3 kb) in length,
wherein the exogenous synthetic spider silk gene is stably integrated in a sixth intron of a fibroin light chain gene, FibL.

16. The transgenic silkworm of claim 15 , wherein the endogenous silkworm promoter is a silkworm-specific U6 promoter.

17. The transgenic silkworm of claim 15 , wherein the exogenous spider silk gene comprises the MaSp1 gene, MaSp2 gene, Ac1 gene, Flag gene, Piri gene or the MiSp1 gene of Nephila clavipes.

18. The transgenic silkworm of claim 15 , wherein the silkworm is Bombyx sp.

19. A transgenic Bombyx silkworm, the silkworm comprising:
a stably-integrated MaSp1 gene or the MiSp1 gene of Nephila clavipes synthetic spider silk gene in a sixth intron of a fibroin light chain gene,
wherein the exogenous spider silk gene is greater than three kilobases (3 kb) in length,
wherein the transgenic silkworm stably expresses the spider silk gene.

20. The transgenic silkworm of claim 19 , wherein the exogenous spider silk gene comprises SEQ ID NO: 25 or SEQ ID NO: 26, or a fragment of either thereof.

Assignees

• Lewis Randolph V
• Xia Lijin
• Zhang Xiaoli
• Jones Justin A
• Univ Utah State

Inventors

• Lewis Randolph V
• Xia Lijin
• Zhang Xiaoli
• Jones Justin A

Classifications

• A01K67/63 Primary

  Genetically modified worms · CPC title

• C07K14/43518

  from spiders · CPC title

• C12N15/90

  Stable introduction of foreign DNA into chromosome · CPC title

• A01K2217/052

  inducing gain of function · CPC title

• C07K14/43586

  from silkworms · CPC title