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Systemic RNAi Delivery to the Muscles of ROSA26 Mice Reduces lacZ Expression

RNAi has potential for therapeutically downregulating the expression of dominantly inherited genes in a variety of human genetic disorders. Here we used the ROSA26 mouse, which constitutively expresses the bacterial lacZ gene in tissues body wide, as a model to test the ability to downregulate gene...

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Detalles Bibliográficos
Autores principales: Wei, Jessica, Chamberlain, Joel R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4134137/
https://www.ncbi.nlm.nih.gov/pubmed/25127128
http://dx.doi.org/10.1371/journal.pone.0102053
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author Wei, Jessica
Chamberlain, Joel R.
author_facet Wei, Jessica
Chamberlain, Joel R.
author_sort Wei, Jessica
collection PubMed
description RNAi has potential for therapeutically downregulating the expression of dominantly inherited genes in a variety of human genetic disorders. Here we used the ROSA26 mouse, which constitutively expresses the bacterial lacZ gene in tissues body wide, as a model to test the ability to downregulate gene expression in striated muscles. Recombinant adeno-associated viral vectors (rAAVs) were generated that express short hairpin RNAs (shRNAs) able to target the lacZ mRNA. Systemic delivery of these rAAV6 vectors led to a decrease of β-galactosidase expression of 30–50-fold in the striated muscles of ROSA26 mice. However, high doses of vectors expressing 21 nucleotide shRNA sequences were associated with significant toxicity in both liver and cardiac muscle. This toxicity was reduced in cardiac muscle using lower vector doses. Furthermore, improved knockdown in the absence of toxicity was obtained by using a shorter (19 nucleotide) shRNA guide sequence. These results support the possibility of using rAAV vectors to deliver RNAi sequences systemically to treat dominantly inherited disorders of striated muscle.
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spelling pubmed-41341372014-08-19 Systemic RNAi Delivery to the Muscles of ROSA26 Mice Reduces lacZ Expression Wei, Jessica Chamberlain, Joel R. PLoS One Research Article RNAi has potential for therapeutically downregulating the expression of dominantly inherited genes in a variety of human genetic disorders. Here we used the ROSA26 mouse, which constitutively expresses the bacterial lacZ gene in tissues body wide, as a model to test the ability to downregulate gene expression in striated muscles. Recombinant adeno-associated viral vectors (rAAVs) were generated that express short hairpin RNAs (shRNAs) able to target the lacZ mRNA. Systemic delivery of these rAAV6 vectors led to a decrease of β-galactosidase expression of 30–50-fold in the striated muscles of ROSA26 mice. However, high doses of vectors expressing 21 nucleotide shRNA sequences were associated with significant toxicity in both liver and cardiac muscle. This toxicity was reduced in cardiac muscle using lower vector doses. Furthermore, improved knockdown in the absence of toxicity was obtained by using a shorter (19 nucleotide) shRNA guide sequence. These results support the possibility of using rAAV vectors to deliver RNAi sequences systemically to treat dominantly inherited disorders of striated muscle. Public Library of Science 2014-08-15 /pmc/articles/PMC4134137/ /pubmed/25127128 http://dx.doi.org/10.1371/journal.pone.0102053 Text en © 2014 Wei, Chamberlain http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Wei, Jessica
Chamberlain, Joel R.
Systemic RNAi Delivery to the Muscles of ROSA26 Mice Reduces lacZ Expression
title Systemic RNAi Delivery to the Muscles of ROSA26 Mice Reduces lacZ Expression
title_full Systemic RNAi Delivery to the Muscles of ROSA26 Mice Reduces lacZ Expression
title_fullStr Systemic RNAi Delivery to the Muscles of ROSA26 Mice Reduces lacZ Expression
title_full_unstemmed Systemic RNAi Delivery to the Muscles of ROSA26 Mice Reduces lacZ Expression
title_short Systemic RNAi Delivery to the Muscles of ROSA26 Mice Reduces lacZ Expression
title_sort systemic rnai delivery to the muscles of rosa26 mice reduces lacz expression
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4134137/
https://www.ncbi.nlm.nih.gov/pubmed/25127128
http://dx.doi.org/10.1371/journal.pone.0102053
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