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Generation of lentiviral transgenic rats expressing Glutamate Receptor Interacting Protein 1 (GRIP1) in brain, spinal cord and testis

Nakagawa, Terunaga and Feliu-Mojer, Monica I. and Wulf, Phebe and Lois, Carlos and Sheng, Morgan and Hoogenraad, Casper C. (2006) Generation of lentiviral transgenic rats expressing Glutamate Receptor Interacting Protein 1 (GRIP1) in brain, spinal cord and testis. Journal of Neuroscience Methods, 152 (1-2). pp. 1-9. ISSN 0165-0270. https://resolver.caltech.edu/CaltechAUTHORS:20180926-143405497

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Abstract

In neuroscience, rats have several advantages over mice as a model organism. For instance, behavioral experiments are more advanced and the larger size of the brain is better suited for surgical manipulation and biochemistry. Furthermore, the vascular physiology of rats is considered closer to human, providing clinical relevance. Because transgenesis rates achieved by conventional pronuclear injection are extremely low (0.2–3.5%), the availability of transgenic rats in neuroscience is limited. Lentivirus infection is an efficient way to integrate exogenous genes into the genome of a one-cell embryo to generate transgenic animals. We report here the generation of synapsin I promoter driven GRIP1-transgenic rats using lentiviral transgenesis. GRIP1 was chosen as a transgene because it interacts with AMPA receptors and is involved in glutamate receptor signaling. From a single infection experiment, 45% of the offspring carried the transgene and 40% achieved germ-line transmission. The expression of GRIP1 was observed at low levels in brain, spinal cord and testis. Interestingly, one transgenic copy lacked a 147 bp fragment in the GRIP1 coding region most likely caused by alternative splicing of genomic lentiviral RNA. Co-immunoprecipitation from rat brains showed that transgenic GRIP1 is in complex with the endogenous GluR2 subunit of AMPA receptors. These results indicate that functional transgenic GRIP1 protein is expressed in rat brain using lentiviral vectors containing a human synapsin I promoter. Tissue specific lentiviral transgenic rats will be a powerful tool for various applications in modern neuroscience.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.jneumeth.2005.08.001DOIArticle
ORCID:
AuthorORCID
Lois, Carlos0000-0002-7305-2317
Additional Information:© 2005 Elsevier B.V. Received 28 June 2005, Revised 23 July 2005, Accepted 3 August 2005, Available online 12 September 2005. We thank Professor Susumu Tonegawa for support and the Department of Comparative Medicine (Bob Marini, Susan Eldman and Alison Hayward) for technical help. C.C.H is recipient of long term fellowship from International Human Frontier Science Program Organization and supported by The Netherlands Organization for Scientific Research (NWO). M.S. is investigator of the Howard Hughes Medical Institute.
Funders:
Funding AgencyGrant Number
Human Frontier Science ProgramUNSPECIFIED
Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO)UNSPECIFIED
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Subject Keywords:Lentiviral vectors; Viral gene transfer; Synapsin promoter; AMPA receptor; GRIP1/ABP; Transgenic rats
Issue or Number:1-2
Record Number:CaltechAUTHORS:20180926-143405497
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20180926-143405497
Official Citation:Terunaga Nakagawa, Monica I. Feliu-Mojer, Phebe Wulf, Carlos Lois, Morgan Sheng, Casper C. Hoogenraad, Generation of lentiviral transgenic rats expressing Glutamate Receptor Interacting Protein 1 (GRIP1) in brain, spinal cord and testis, Journal of Neuroscience Methods, Volume 152, Issues 1–2, 2006, Pages 1-9, ISSN 0165-0270, https://doi.org/10.1016/j.jneumeth.2005.08.001. (http://www.sciencedirect.com/science/article/pii/S0165027005002797)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:89974
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:26 Sep 2018 21:46
Last Modified:03 Oct 2019 20:21

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