Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Novel shRNA for increasing animal fertility and application thereof

A new type of animal technology, applied in the fields of application, plant genetic improvement, botanical equipment and methods, etc., can solve the problems of inability to transfer high-breeding traits of parents to offspring animals, etc.

Inactive Publication Date: 2012-04-11
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
View PDF2 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] Inhibin active immunization works only on the parent animals, and cannot pass the high-breeding traits of the parents to the offspring animals

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Novel shRNA for increasing animal fertility and application thereof
  • Novel shRNA for increasing animal fertility and application thereof
  • Novel shRNA for increasing animal fertility and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Construction and identification of ovine inhibin shRNA lentiviral interference vector

[0033] 1. Research methods

[0034] 1. Construction of shRNA interference vector

[0035] Four shRNA targets were screened against the cDNA sequence of the sheep INHA gene (L28815.1) using the online shRNA design tool of Invitrogen Company http: / / rnaidesigner.invitrogen.com / rnaiexpress / . The DNA sequence of the target site is shown in the sequence table Sequnence NO.1, Sequnence NO.2, Sequnence NO.3, Sequnence NO.4. Take each target sequence as the 5' end, its reverse complementary sequence as the 3' end, connect the 5' end and the 3' end with a stem-loop structure sequence, and then introduce BamHI and SalI at the 5' end and 3' end respectively Restriction site, which constitutes the coding (template) sequence of the shRNA. Then, a sequence with no significant similarity (homology) to all known gene coding sequences in sheep was designed as a negative control sequence, ...

Embodiment 2

[0117] Embodiment 2 gonad injection method prepares transgenic sheep

[0118] 1. Materials and methods

[0119] 1. Materials:

[0120] 1) Experimental animals

[0121] The experimental small-tailed Han sheep were purchased from the small-tailed Han sheep breeding base in Yuncheng, Shandong, including 46 ewes (12-18 months old) and 10 rams (12-20 months old). Before starting this experiment, the fecundity-related gene FecB gene of rams and ewes had been identified, and the obtained identification results showed that 29 of the 46 ewes were heterozygous for FecB, and 11 did not contain the FecB gene. The FecB genotypes of other sheep were not detected; 4 of the 10 rams were recessive homozygous for FecB, and 6 were heterozygous for FecB.

[0122] 2) Main reagents

[0123]

[0124]

[0125] 3) Main equipment

[0126] Surgical instruments (scalpel, surgical needle, surgical thread, wound towel, medical gauze, hemostatic forceps, needle-holding forceps, surgical scissors,...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses novel shRNA for increasing animal fertility and an application thereof, and the shRNA is characterized in that the shRNA comprises gene sequences of Sequence NO. 15 and Sequence NO. 16. According to the invention, several pairs of shRNA single-chain oligonucleotides are designed and synthesized based on the nucleotide sequence of sheep interferon INHA gene alpha subunit; an INHA shRNA interference vector pGFP-V-RS-shRNA is constructed; the interference effect is screened by a RT-PCR method so as to obtain a pGFP-V-RS-shRNA vector with good inhibition effect; the vector comprises gene sequences of Sequence NO. 15 and Sequence NO. 16; and a shRNA lentivirus interference vector pLenti6-shRNA3 is further constructed which comprises the gene sequences of Sequence NO. 15 and Sequence NO. 16. The invention also discloses an application of the novel vector.

Description

technical field [0001] The invention relates to the construction of a novel carrier. Specifically, the present invention designs and synthesizes shRNA single-stranded oligonucleotides for the nucleotide sequence of the α subunit of the sheep interferon INHA gene, constructs shRNA lentiviral interference vectors, and obtains new vectors with good application prospects. Background technique [0002] Inhibin (INH) is a peptide gonadotropin that can regulate the secretion of follicle-stimulating hormone (FSH) through pituitary negative feedback. It is a member of the transforming growth factor β (TGF-β) superfamily, mainly composed of ovarian granulosa cells of female animals, Sertoli cells and Leydig cells of male animals [Hayes FJ, Hall JE, Boepple PA, Crowley WF, Jr .Clinical review 96: Differential control of gonadotropin secretion in the human: endocrine role of inhibin. J Clin Endocrinol Metab, 1998, 83(6): 1835-41.] Secreted and widely distributed in multiple organs of t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/867A01K67/027
Inventor 苗向阳张瑞杰
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com