I-SceI meganuclease-mediated transgenesis in Xenopus
Article first published online: 28 OCT 2005
Copyright © 2005 Wiley-Liss, Inc.
Volume 235, Issue 1, pages 247–252, January 2006
How to Cite
Pan, F. C., Chen, Y., Loeber, J., Henningfeld, K. and Pieler, T. (2006), I-SceI meganuclease-mediated transgenesis in Xenopus. Dev. Dyn., 235: 247–252. doi: 10.1002/dvdy.20608
- Issue published online: 7 DEC 2005
- Article first published online: 28 OCT 2005
- Manuscript Accepted: 1 SEP 2005
- Manuscript Revised: 27 JUN 2005
- Manuscript Received: 4 APR 2005
- Deutsche Forschungsgemeinschaft. Grant Number: PI 159/8-1
- I-SceI meganuclease;
Several experimental approaches have been described to generate transgenic frogs. Here, we report on the application of a novel method in Xenopus, making use of I-SceI meganuclease. The characteristic feature of this endonuclease is that it has an extended recognition site of 18 bp, which is expected to exist only once in 7 × 1010 bp of random DNA sequences. Various reporter constructs flanked by two I-SceI recognition sites were injected together with the I-SceI meganuclease into one-cell stage Xenopus embryos. We observed an overall transgenesis frequency of 10% or more under optimized condition. The injected genes were integrated into the genome and transmitted to F1 offspring. Southern blot analysis showed that between one and eight copies of the transgene were integrated. Meganuclease-aided transgenesis, thus, provides a simple and highly efficient tool for transgenesis in Xenopus. Developmental Dynamics 235:247–252, 2006. © 2005 Wiley-Liss, Inc.