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Transgenic animals with customizable traits

A transgenic animal, purpose technology, applied in animal cells, genetic engineering, biochemical equipment and methods, etc.

Inactive Publication Date: 2016-02-24
AGGENETICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Transgenic animals with customizable traits
  • Transgenic animals with customizable traits
  • Transgenic animals with customizable traits

Examples

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preparation example Construction

[0201] Preparation method of non-human transgenic animal

[0202] A variety of arbitrary methods can be used to introduce transgenes into non-human animals to create transgenic animals. Such techniques are well known in the art and include, but are not limited to, pronuclear microinjection, virus injection, and transformation of embryonic stem cells and iPS cells. Methods that can be used to create transgenic animals include, but are not limited to, those described in J.P. Sundberg and T. Ichiki, Eds., Genetically Engineered Mice Handbook, CRC Press; 2006; M.H. Hofker and I. van Deursen, Eds., Transgenic Mouse Methods and Protocols, Humana Press, 2002; A.L. Joyner, GeneTargeting: APracticalApproach,OxfordUniversityPress,2000;ManipulatingtheMouseEmbryo:ALaboratoryManual,第三版,ColdSpringHarborLaboratoryPress;2002,ISBN-10:0879695919;K.Turksen(Ed.),Embryonicstemcells:methodsandprotocolsinMethodsMol.Biol.2002;185,HumanaPress;CurrentProtocolsinStemCellBiology,ISBN:978047015180; Metho...

Embodiment 1

[0224] Example 1 - Customization of Animal Skin or Fur Coloring

[0225] This example provides an embodiment of a genetic construct for customizing animal skin or fur coloration

[0226] Animals with custom skin or fur coloring or patterns

[0227] C3H / HeJ mice with brown ("Agouti" stained) skin color were transgenic to express the following three constructs: 1) a construct comprising a keratin-14 specific promoter, a loxp cassette (which includes a nucleic acid encoding a red fluorescent protein ), a pigment cassette (which includes a nucleic acid encoding a dominant black (ΔG23) beta defensin 103 protein) and (with introns) SV40 polyadenylation sequence; 2) a construct comprising an encoding reverse tetracycline trans A nucleic acid molecule of the formula activator (rtTA) and an SV40 polyadenylation sequence, said nucleic acid molecule being operably linked to a keratin-14 specific promoter that initiates transcription of the nucleic acid encoding rtTA; and 3 ) construct ...

Embodiment 2

[0236] Example 2 - Mice with Custom Fur Colors and Patterns

[0237] This example shows the creation of genetically modified mice whose fur color can be permanently altered by transdermal administration of HTNCre recombinase, which readily crosses cell membranes.

[0238] Figure 4 Genetic constructs used to create custom patterns and colors in mouse skin or fur are shown. The construct contains the keratin 14 promoter, which is expressed in all skin fibroblasts to induce a dominant black form of the signaling molecule β-defensin 103. Expression of β-defensin 103 was blocked by loxP site cleavage nucleic acid encoding ring finger protein (RFP), which was used as a marker.

[0239] Agouti mice were transgenic to express a transgene encoding the "dominant black" signaling molecule β-defensin 103, and transgene expression was activated by administration of recombinase.

[0240] Figure 5A and B are showing two transgenic mice in which expression of dominant melanin protein wa...

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Abstract

Disclosed are materials and methods for creating customizable traits in animals. In the demonstration of the principle of the subject invention, a keratin-14 specific promoter is used with red fluorescent protein in the loxp cassette, dominant black ([Delta]G23) beta defensin 103 in the pigment cassette, and an SV40 (with intron) polyadenylation sequence. When Cre recombinase (or HTNCre) is applied to the animal's skin in a carrier base (e.g., lipid bilayers), fur is permanently genetically modified to tum black in the shape in which theHTNCre was applied.

Description

[0001] Cross References to Related Applications [0002] This application is a continuation-in-part of U.S. Patent Application No. 13 / 837,405 filed March 15, 2013, which is a continuation-in-part of U.S. Patent Application No. 13 / 837,405 filed on February 15, 2013 A continuation-in-part of application 13 / 768,760, U.S. Patent Application No. 13 / 768,760 claims priority to U.S. Patent Application No. 61 / 598,987, filed February 15, 2012, the entire contents of which are incorporated by reference way incorporated into this article. [0003] The sequence listing for this application is labeled SeqList-14Marl3_ST25.txt, created on March 14, 2013 and is 85KB in size. The entire contents of the Sequence Listing are incorporated herein by reference. technical field [0004] The present invention relates to the genetic modification of animals to produce controlled changes in specific characteristics, for example, changes in skin and / or fur colouration. Background technique [0005] ...

Claims

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Application Information

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IPC IPC(8): A01K67/027C12N5/07C12N15/09
CPCA01K67/0275A01K2227/101A01K2267/02C12N15/8509
Inventor 詹姆士·韦斯特
Owner AGGENETICS
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