Targeted gene deletion of miRNAs in mice by TALEN system

Mice are among the most valuable model animal species with an enormous amount of heritage in genetic modification studies. However, targeting genes in mice is sometimes difficult, especially for small genes, such as microRNAs (miRNAs) and targeting genes in repeat sequences. Here we optimized the ap...

Full description

Saved in:
Bibliographic Details
Published in:PloS one 2013-10, Vol.8 (10), p.e76004
Main Authors: Takada, Shuji, Sato, Tempei, Ito, Yoshiaki, Yamashita, Satoshi, Kato, Tomoko, Kawasumi, Miyuri, Kanai-Azuma, Masami, Igarashi, Arisa, Kato, Tomomi, Tamano, Moe, Asahara, Hiroshi
Format: Article
Language:English
Subjects:
Animal species
Animals
Base Sequence
Childrens health
Chromosomes
Clonal deletion
Deoxyribonucleases, Type II Site-Specific - genetics
Deoxyribonucleases, Type II Site-Specific - metabolism
Deoxyribonucleic acid
DNA
DNA, Intergenic - genetics
DNA, Intergenic - metabolism
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Embryo, Mammalian
Female
Gene Deletion
Gene mutation
Gene sequencing
Gene targeting
Gene Targeting - methods
Genes
Genetic Engineering
Genetic modification
Genetically modified organisms
Genomes
House mouse
Mice
Mice, Inbred ICR
Microinjection
Microinjections
MicroRNA
MicroRNAs
MicroRNAs - genetics
MicroRNAs - metabolism
miRNA
Molecular Sequence Data
Oocytes
Oocytes - cytology
Oocytes - growth & development
Oocytes - metabolism
Plasmids
Poly(A)
Polyadenine
Polyadenylation
Pseudopregnancy
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Ribonucleic acid
RNA
Rodents
Stem cells
Transcription activator-like effector nucleases
Xenopus
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Mice are among the most valuable model animal species with an enormous amount of heritage in genetic modification studies. However, targeting genes in mice is sometimes difficult, especially for small genes, such as microRNAs (miRNAs) and targeting genes in repeat sequences. Here we optimized the application of TALEN system for mice and successfully obtained gene targeting technique in mice for intergenic region and series of microRNAs. Microinjection of synthesized RNA of TALEN targeting each gene in one cell stage of embryo was carried out and injected oocytes were transferred into pseudopregnant ICR female mice, producing a high success rate of the targeted deletion of miRNA genes. In our condition, TALEN RNA without poly(A) tail worked better than that of with poly(A) tail. This mutated allele in miRNA was transmitted to the next generation, suggesting the successful germ line transmission of this targeting method. Consistent with our notion of miRNAs maturation mechanism, in homozygous mutant mice of miR-10a, the non- mutated strand of miRNAs expression was completely diminished. This method will lead us to expand and accelerate our genetic research using mice in a high throughput way.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0076004