I-SceI-Mediated Transgenesis in Xenopus

Anna Noble; Anita Abu-Daya; Matthew J. Guille

doi:10.1101/pdb.prot107011

I-SceI-Mediated Transgenesis in Xenopus

Anna Noble
, Anita Abu-Daya
, Matthew J. Guille^*

^*Corresponding author for this work

Institute of Life Sciences and Healthcare

Research output: Contribution to journal › Article › peer-review

Abstract

Transgenic frogs can be very efficiently generated using I-SceI meganuclease, a nuclease with an 18-bp recognition site. The desired transgene must be flanked by I-SceI sites, in either a plasmid or a polymerase chain reaction (PCR) product. After a short in vitro digestion with the meganuclease, the complete reaction is injected into fertilized eggs, where the enzyme mediates genomic integration by an unknown mechanism. Posttransgenesis development is typically normal, and up to 70% of the embryos integrate the transgene.

Original language	English
Pages (from-to)	1-6
Number of pages	6
Journal	Cold Spring Harbor Protocols
Volume	2022
Issue number	6
Early online date	8 Feb 2022
DOIs	https://doi.org/10.1101/pdb.prot107011
Publication status	Published - 1 Jun 2022

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title = "I-SceI-Mediated Transgenesis in Xenopus",

abstract = "Transgenic frogs can be very efficiently generated using I-SceI meganuclease, a nuclease with an 18-bp recognition site. The desired transgene must be flanked by I-SceI sites, in either a plasmid or a polymerase chain reaction (PCR) product. After a short in vitro digestion with the meganuclease, the complete reaction is injected into fertilized eggs, where the enzyme mediates genomic integration by an unknown mechanism. Posttransgenesis development is typically normal, and up to 70\% of the embryos integrate the transgene.",

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AB - Transgenic frogs can be very efficiently generated using I-SceI meganuclease, a nuclease with an 18-bp recognition site. The desired transgene must be flanked by I-SceI sites, in either a plasmid or a polymerase chain reaction (PCR) product. After a short in vitro digestion with the meganuclease, the complete reaction is injected into fertilized eggs, where the enzyme mediates genomic integration by an unknown mechanism. Posttransgenesis development is typically normal, and up to 70% of the embryos integrate the transgene.

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I-SceI-Mediated Transgenesis in Xenopus

Abstract

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