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Method for down-regulating zebra fish lmna gene by using morpholino and established progeria disease model

A disease model, the technology of lmna-egfp-pcs2, is applied in the field of genetic engineering to achieve the effect of easy establishment of disease model, fast maturation and small size

Inactive Publication Date: 2016-11-16
GUIZHOU MEDICAL UNIV
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Problems solved by technology

by establishing lmna The zebrafish model with down-regulated genes can provide an animal model for the study of human laminopathy, but there is no ideal technical solution to solve this technical problem in this field

Method used

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  • Method for down-regulating zebra fish lmna gene by using morpholino and established progeria disease model
  • Method for down-regulating zebra fish lmna gene by using morpholino and established progeria disease model
  • Method for down-regulating zebra fish lmna gene by using morpholino and established progeria disease model

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Embodiment 1

[0024] 1. Materials and methods

[0025] 1. Morpholino oligonucleotide sequence design

[0026] Inhibition of endogenous lmna gene expression. Morpholino oligonucleotides are designed for the gene coding start site ATG, and act on the 5' non-translation region (untranslation region, UTR) region near the translation start site AUG of mRNA or upstream of AUG, and its combination with mRNA It hinders the movement of the translation initiation complex of the ribosome to the translation initiation site, resulting in the inability to initiate the synthesis process of the polypeptide. Since ATG for lmna The initiation of gene translation plays a vital role, so when designing morpholino oligonucleotides, avoid the ATG initiation region, and design antisense oligonucleotides in its 5'UTR region to enable translation of exogenous mRNA Expressions are not affected, as in figure 1 . lmna -MO's target sequence is located at lmna The 5'UTR end of the gene, the sequence of control-M...

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Abstract

The invention discloses a method for down-regulating a zebra fish lmna gene by using morpholino and an established progeria disease model; a morpholino oligonucleotide recombinant plasmid lmna-MO aiming at the zebra fish lmna gene and a lmna-EGFP-pCS<2+> recombinant plasmid for specific instruction of the expression quantity of the lmna gene are respectively designed and constructed; the two recombinant plasmids are co-injected into zebra fish embryos by a microinjection way, and the expression quantity of green fluorescent proteins is used as a lmna expression down-regulation mark, so that the zebra fish animal model with down-regulated lmna gene expression is established. The zebra fish animal model with down-regulated lmna gene expression is established, and can provide an animal model for deep research of laminopathy.

Description

technical field [0001] The invention relates to a method for establishing a disease model by down-regulating the zebrafish lmna gene and a premature aging disease model established by the method, belonging to the technical field of genetic engineering. Background technique [0002] Lamins can maintain the stability of the nucleus, connect the nucleus and the cytoskeleton, and regulate nuclear chromatin organization and gene expression. Mutations in lamins can lead to a series of disorders, including premature aging, muscular dystrophy and dilated cardiomyopathy, collectively referred to as laminopathy. [0003] Lamins (lamins) are members of the V-type intermediate fibrin family (intermediate filament, IF). Mutations of Lamins and their related proteins can lead to human laminopathy. Since LaminsA / C is expressed in almost all cells and tissues, the high degree of tissue specificity and the prevalence of diseases caused by different mutation sites in the same gene are relati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N15/65A01K67/027A61K49/00
CPCC12N15/8509A01K67/0275A01K2217/05A01K2227/40A01K2267/0306A61K49/0008C12N15/65C12N2800/106
Inventor 舒莉萍何志旭黄惠敏叶芝旭王珺侯仕芳李雪华
Owner GUIZHOU MEDICAL UNIV
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