Rapid reverse genetics systems for N. furzeri, a suitable model organism to study vertebrate aging
Year: 2022
Type: preprint
Abstract: The African turquoise killifish (N. furzeri) is a useful model organism for studying aging, age-related diseases, and embryonic diapause. CRISPR/Cas9-mediated gene knockout and Tol2 transposon-mediated transgenesis in N. furzeri have been reported previously. However, these methods take time to generate knockout and transgenic fish. In addition, knock-in technology that inserts large DNA fragments as fluorescent reporter constructs into the target gene in N. furzeri has not yet been established. Here, we show that triple-target CRISPR-mediated single gene disruption efficiently produces whole-body biallelic knockout and enables the examination of gene function in the F0 generation. In addition, we developed a method for creating the knock-in reporter N. furzeri without crossing by optimizing the CRISPR/Cas9 system. These methods drastically reduce the experimental duration and we think that these will accelerate aging and developmental studies using N. furzeri.
Source: Research Square
Authors: Masayuki Oginuma, Moana Nishida, Tomomi Ohmura-Adachi, Kota Abe, Shohei Ogamino, Chihiro Mogi, Hideaki Matsui, Tohru Ishitani
Institutions: Gunma University, Niigata University, The University of Osaka
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Awards: G1878932742, 超短命魚で解き明かすモルフォゲン制御による個体老化機構, G4055240873, G5503133928, Analysis of metabolic pathways that control organismal aging using the short-lived vertebrate model., Gradients of energy metabolism: A novel player involved on the vertebrate embryo patterning, Molecular Basis underlying morphogen gradient-mediated tissue homeostatic regulation, 老化機構・制御研究拠点, G8735883516
OpenAlex ID: W4282018020