The Ars insulator facilitates I-SceI meganuclease-mediated transgenesis in the sea urchin embryo

Developmental Dynamics : an Official Publication of the American Association of Anatomists
Hiroshi OchiaiTakashi Yamamoto

Abstract

For the efficient generation of transgenic sea urchins, we have adopted an I-SceI meganuclease-mediated transgenesis method. Several types of promoter-GFP gene constructs flanked by two I-SceI recognition sequences were co-injected with I-SceI into sea urchin fertilized eggs. Using cell-lineage-specific promoter constructs, the frequency of transgene expression was elevated, and their level of mozaicism was reduced. The addition of the Ars insulator sequence, which is known to block the enhancer activity and protect transgenes from position effects, led to a reduction in ectopic transgene expression and an elevation of transgene expression frequency in this I-SceI-mediated system. However, the magnitude of the effects of the Ars insulator was dependent upon the promoter constructs. QPCR analysis also showed that the Ars insulator increases the transgene copy number. These results suggest that the I-SceI-mediated method using the Ars insulator is advantageous for transgenesis in the sea urchin embryo.

References

Sep 1, 1988·Developmental Biology·B R Hough-EvansE H Davidson
Aug 1, 1983·Archives of Biochemistry and Biophysics·K Okabayashi, E Nakano
Sep 3, 1996·Proceedings of the National Academy of Sciences of the United States of America·D A GdulaV G Corces
Jan 25, 2000·Developmental Biology·L M Angerer, R C Angerer
Sep 28, 2002·Mechanisms of Development·Violette ThermesJean-Stéphane Joly
Apr 18, 2003·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Izumi Oda-Ishii, Hidetoshi Saiga
Apr 30, 2003·Genesis : the Journal of Genetics and Development·Karine DeschetWilliam C Smith
Jul 21, 2004·Current Opinion in Genetics & Development·Paola Oliveri, Eric H Davidson
Dec 18, 2004·Methods in Cell Biology·Clemens GrabherJoachim Wittbrodt
Mar 30, 2005·Proceedings of the National Academy of Sciences of the United States of America·Tetsuya KoideKen W Y Cho
Nov 1, 2005·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Fong Cheng PanTomas Pieler
May 27, 2006·Science·Kaoru S ImaiYutaka Satou
Aug 23, 2006·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Satoshi WatanabeKoji Akasaka
Mar 21, 2007·Development Genes and Evolution·Mamiko YajimaKoji Akasaka
May 10, 2007·Nature Protocols·Hajime OginoRobert M Grainger
Aug 3, 2007·Nucleic Acids Research·Anat Haviv-ChesnerMartin Kupiec
Dec 6, 2007·Genome Biology·Clemens Grabher, Joachim Wittbrodt

❮ Previous
Next ❯

Citations

May 19, 2010·BMC Biotechnology·Jean-Charles GabillardPierre-Yves Rescan
Jul 8, 2010·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Hiroshi OchiaiTakashi Yamamoto
Nov 10, 2018·Developmental Biology·Paul J MinorChristopher J Lowe

❮ Previous
Next ❯

Related Concepts

Related Feeds

CREs: Gene & Cell Therapy

Gene and cell therapy advances have shown promising outcomes for several diseases. The role of cis-regulatory elements (CREs) is crucial in the design of gene therapy vectors. Here is the latest research on CREs in gene and cell therapy.

Related Papers

Proceedings of the National Academy of Sciences of the United States of America
Eduard RenferUlrich Technau
Developmental Dynamics : an Official Publication of the American Association of Anatomists
Fong Cheng PanTomas Pieler
Mechanisms of Development
Violette ThermesJean-Stéphane Joly
© 2022 Meta ULC. All rights reserved