A modified TALEN-based strategy for rapidly and efficiently generating knockout mice for kidney development studies

The transcription activator-like effector nucleases (TALENs) strategy has been widely used to delete and mutate genes in vitro. This strategy has begun to be used for in vivo systemic gene manipulation, but not in an organ-specific manner. In this study, we developed a modified, highly efficient TAL...

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Bibliographic Details
Published in:PloS one 2014-01, Vol.9 (1), p.e84893-e84893
Main Authors: Liu, Yunhong, Lv, Xiaoyan, Tan, Ruizhi, Liu, Tianming, Chen, Tielin, Li, Mi, Liu, Yuhang, Nie, Fang, Wang, Xiaoyan, Zhou, Puhui, Chen, Mianzhi, Zhou, Qin
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Biology
Cell Nucleus - genetics
Cloning
Deoxyribonucleic acid
Design
DNA
DNA binding proteins
DNA Restriction Enzymes - metabolism
Efficiency
Fluorescence
Frameshift Mutation
Gene Knockout Techniques - methods
Gene manipulation
Gene mutation
Genes, Reporter - genetics
Genetic aspects
Genetic engineering
Genomes
Genotypes
HEK293 Cells
Homologous Recombination - genetics
Homozygote
Hospitals
Humans
Kidney - cytology
Kidney - growth & development
Kidney - metabolism
Kidney - surgery
Kidneys
Laboratory animals
Medical schools
Medicine
Mice
Mutagenesis
Mutation
Nephrectomy
Nephrology
Nuclease
Nucleases
Offspring
Phenotype
Plasmids
Rodents
Strategy
Time Factors
Transcription
Transcription activator-like effector nucleases
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Summary:The transcription activator-like effector nucleases (TALENs) strategy has been widely used to delete and mutate genes in vitro. This strategy has begun to be used for in vivo systemic gene manipulation, but not in an organ-specific manner. In this study, we developed a modified, highly efficient TALEN strategy using a dual-fluorescence reporter. We used this modified strategy and, within 5 weeks, we successfully generated kidney proximal tubule-specific gene Ttc36 homozygous knockout mice. Unilateral nephrectomy was performed on the 6-week-old founders (F0) to identify the knockout genotype prior to the birth of the offspring. This strategy was found to have little effect on reproduction in the knockout mice and inheritability of the knockout genotypes. The modified TALEN knockout strategy in combination with unilateral nephrectomy can be readily used for studies of gene function in kidney development and diseases.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0084893