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Characterization of knockin mice at the Rosa26, Tac1 and Plekhg1 loci generated by homologous recombination in oocytes

Design and engineering of complex knockin mice has revolutionized the in vivo manipulation of genetically defined cells. Recently development of the bacterial clustered regularly interspersed short palindromic repeats (CRISPR) associated protein 9 (Cas9) system for single site cleavage of mammalian...

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Autores principales: Wu, Youmei, Luna, María José, Bonilla, Lauren S., Ryba, Nicholas J. P., Pickel, James M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5828354/
https://www.ncbi.nlm.nih.gov/pubmed/29485996
http://dx.doi.org/10.1371/journal.pone.0193129
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author Wu, Youmei
Luna, María José
Bonilla, Lauren S.
Ryba, Nicholas J. P.
Pickel, James M.
author_facet Wu, Youmei
Luna, María José
Bonilla, Lauren S.
Ryba, Nicholas J. P.
Pickel, James M.
author_sort Wu, Youmei
collection PubMed
description Design and engineering of complex knockin mice has revolutionized the in vivo manipulation of genetically defined cells. Recently development of the bacterial clustered regularly interspersed short palindromic repeats (CRISPR) associated protein 9 (Cas9) system for single site cleavage of mammalian genomes has opened the way for rapid generation of knockin mice by targeting homology directed repair to selected cleavage sites. We used this approach to generate new lines of mice that will be useful for a variety of aspects of neuroscience research. These lines have been bred to homozygosity and details of the expression and function of the transgenes are reported. Two lines target the Rosa26-locus and have been engineered to allow Cre-dependent expression of the avian tva receptor, and Cre-dependent expression of a cell surface targeted spaghetti-monster carrying many copies of the “ollas-tag”. Another line expresses red fluorescent protein and tva in Tac1-positive neurons; the fourth line targets FlpO expression to Plekhg1 expressing neurons, providing a powerful approach to modify gene expression in thalamic excitatory neurons.
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spelling pubmed-58283542018-03-19 Characterization of knockin mice at the Rosa26, Tac1 and Plekhg1 loci generated by homologous recombination in oocytes Wu, Youmei Luna, María José Bonilla, Lauren S. Ryba, Nicholas J. P. Pickel, James M. PLoS One Research Article Design and engineering of complex knockin mice has revolutionized the in vivo manipulation of genetically defined cells. Recently development of the bacterial clustered regularly interspersed short palindromic repeats (CRISPR) associated protein 9 (Cas9) system for single site cleavage of mammalian genomes has opened the way for rapid generation of knockin mice by targeting homology directed repair to selected cleavage sites. We used this approach to generate new lines of mice that will be useful for a variety of aspects of neuroscience research. These lines have been bred to homozygosity and details of the expression and function of the transgenes are reported. Two lines target the Rosa26-locus and have been engineered to allow Cre-dependent expression of the avian tva receptor, and Cre-dependent expression of a cell surface targeted spaghetti-monster carrying many copies of the “ollas-tag”. Another line expresses red fluorescent protein and tva in Tac1-positive neurons; the fourth line targets FlpO expression to Plekhg1 expressing neurons, providing a powerful approach to modify gene expression in thalamic excitatory neurons. Public Library of Science 2018-02-27 /pmc/articles/PMC5828354/ /pubmed/29485996 http://dx.doi.org/10.1371/journal.pone.0193129 Text en https://creativecommons.org/publicdomain/zero/1.0/ This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 (https://creativecommons.org/publicdomain/zero/1.0/) public domain dedication.
spellingShingle Research Article
Wu, Youmei
Luna, María José
Bonilla, Lauren S.
Ryba, Nicholas J. P.
Pickel, James M.
Characterization of knockin mice at the Rosa26, Tac1 and Plekhg1 loci generated by homologous recombination in oocytes
title Characterization of knockin mice at the Rosa26, Tac1 and Plekhg1 loci generated by homologous recombination in oocytes
title_full Characterization of knockin mice at the Rosa26, Tac1 and Plekhg1 loci generated by homologous recombination in oocytes
title_fullStr Characterization of knockin mice at the Rosa26, Tac1 and Plekhg1 loci generated by homologous recombination in oocytes
title_full_unstemmed Characterization of knockin mice at the Rosa26, Tac1 and Plekhg1 loci generated by homologous recombination in oocytes
title_short Characterization of knockin mice at the Rosa26, Tac1 and Plekhg1 loci generated by homologous recombination in oocytes
title_sort characterization of knockin mice at the rosa26, tac1 and plekhg1 loci generated by homologous recombination in oocytes
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5828354/
https://www.ncbi.nlm.nih.gov/pubmed/29485996
http://dx.doi.org/10.1371/journal.pone.0193129
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