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Title: Direct gene disruption by TALENs in medaka embryos
Authors: Wang, T.
Hong, Y. 
Keywords: Dnd
Gene disruption
Gene targeting
Genome editing
Issue Date: 10-Jun-2014
Citation: Wang, T., Hong, Y. (2014-06-10). Direct gene disruption by TALENs in medaka embryos. Gene 543 (1) : 28-33. ScholarBank@NUS Repository.
Abstract: Targeted gene disruption (GD) is powerful for generating genetic alterations in animal genomes. Engineered endonucleases such as zinc finger nucleases and transcription activator-like effector nucleases (TALENs) allow for GD directly in animal embryos to achieve germline transmission. Here we report procedures and parameters of TALEN-mediated GD in the fish medaka by using a germ cell-specific gene dnd as a model. Embryos at the 1-cell stage were microinjected with synthetic TALEN mRNAs and examined for the survival rate and GD efficiency. Medaka embryos can tolerate a high dosage of TALEN-mRNA injection and exhibit a steadily increasing GD efficiency with increasing mRNA dosages before peaking at 100ng/μl. This dosage produced ~24% efficiency for somatic GD. Some of the animals from manipulated embryos developed into fertile female and male. Most importantly, four fish (3 males and 1 female) examined by progeny-test were able to produce GD-bearing male and female gametes for germline transmission to F1 generation at ~10% efficiency. Therefore, TALEN is proficient for somatic and germline GD in medaka embryos, and disruption of one dnd copy does not compromise somatic development and gamete production. © 2014 Elsevier B.V.
Source Title: Gene
ISSN: 18790038
DOI: 10.1016/j.gene.2014.04.013
Appears in Collections:Staff Publications

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