Cargando…

An efficient rAAV vector for protein expression in cortical parvalbumin expressing interneurons

Recombinant adeno—associated viruses (rAAV) are extensively used in both research and clinical applications. Despite significant advances, there is a lack of short promoters able to drive the expression of virus delivered genes in specific classes of neurons. We designed an efficient rAAV vector sui...

Descripción completa

Detalles Bibliográficos
Autores principales: Tkatch, Tatiana, Rysevaite-Kyguoliene, Kristina, Sabeckis, Ignas, Sabeckiene, Deimante, Pauza, Dainius H., Baranauskas, Gytis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9596399/
https://www.ncbi.nlm.nih.gov/pubmed/36284123
http://dx.doi.org/10.1038/s41598-022-21867-0
_version_ 1784815862344056832
author Tkatch, Tatiana
Rysevaite-Kyguoliene, Kristina
Sabeckis, Ignas
Sabeckiene, Deimante
Pauza, Dainius H.
Baranauskas, Gytis
author_facet Tkatch, Tatiana
Rysevaite-Kyguoliene, Kristina
Sabeckis, Ignas
Sabeckiene, Deimante
Pauza, Dainius H.
Baranauskas, Gytis
author_sort Tkatch, Tatiana
collection PubMed
description Recombinant adeno—associated viruses (rAAV) are extensively used in both research and clinical applications. Despite significant advances, there is a lack of short promoters able to drive the expression of virus delivered genes in specific classes of neurons. We designed an efficient rAAV vector suitable for the rAAV-mediated gene expression in cortical interneurons, mainly in the parvalbumin expressing cells. The vector includes a short parvalbumin promoter and a specialized poly(A) sequence. The degree of conservation of the parvalbumin gene adjoining non-coding regions was used in both the promoter design and the selection of the poly(A) sequence. The specificity was established by co-localizing the fluorescence of the virus delivered eGFP and the antibody for a neuronal marker. rAAV particles were injected in the visual cortex area V1/V2 of adult rats (2–4 months old). Neurons expressing the virus delivered eGFP were mainly positive for interneuronal markers: 66.5 ± 2.8% for parvalbumin, 14.6 ± 2.4% for somatostatin, 7.1 ± 1.2% for vasoactive intestinal peptide, 2.8 ± 0.6% for cholecystokinin. Meanwhile, only 2.1 ± 0.5% were positive for CaMKII, a marker for principal cells in the cortex. The efficiency of the construct was verified by optogenetic experiments: the expression of the virus delivered ChR2 channels was sufficient to evoke by blue light laser high frequency bursts of action potentials in putative fast spiking neurons. We conclude that our promoter allows highly specific expression of the rAAV delivered cDNAs in cortical interneurons with a strong preference for the parvalbumin positive cells.
format Online
Article
Text
id pubmed-9596399
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-95963992022-10-27 An efficient rAAV vector for protein expression in cortical parvalbumin expressing interneurons Tkatch, Tatiana Rysevaite-Kyguoliene, Kristina Sabeckis, Ignas Sabeckiene, Deimante Pauza, Dainius H. Baranauskas, Gytis Sci Rep Article Recombinant adeno—associated viruses (rAAV) are extensively used in both research and clinical applications. Despite significant advances, there is a lack of short promoters able to drive the expression of virus delivered genes in specific classes of neurons. We designed an efficient rAAV vector suitable for the rAAV-mediated gene expression in cortical interneurons, mainly in the parvalbumin expressing cells. The vector includes a short parvalbumin promoter and a specialized poly(A) sequence. The degree of conservation of the parvalbumin gene adjoining non-coding regions was used in both the promoter design and the selection of the poly(A) sequence. The specificity was established by co-localizing the fluorescence of the virus delivered eGFP and the antibody for a neuronal marker. rAAV particles were injected in the visual cortex area V1/V2 of adult rats (2–4 months old). Neurons expressing the virus delivered eGFP were mainly positive for interneuronal markers: 66.5 ± 2.8% for parvalbumin, 14.6 ± 2.4% for somatostatin, 7.1 ± 1.2% for vasoactive intestinal peptide, 2.8 ± 0.6% for cholecystokinin. Meanwhile, only 2.1 ± 0.5% were positive for CaMKII, a marker for principal cells in the cortex. The efficiency of the construct was verified by optogenetic experiments: the expression of the virus delivered ChR2 channels was sufficient to evoke by blue light laser high frequency bursts of action potentials in putative fast spiking neurons. We conclude that our promoter allows highly specific expression of the rAAV delivered cDNAs in cortical interneurons with a strong preference for the parvalbumin positive cells. Nature Publishing Group UK 2022-10-25 /pmc/articles/PMC9596399/ /pubmed/36284123 http://dx.doi.org/10.1038/s41598-022-21867-0 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Tkatch, Tatiana
Rysevaite-Kyguoliene, Kristina
Sabeckis, Ignas
Sabeckiene, Deimante
Pauza, Dainius H.
Baranauskas, Gytis
An efficient rAAV vector for protein expression in cortical parvalbumin expressing interneurons
title An efficient rAAV vector for protein expression in cortical parvalbumin expressing interneurons
title_full An efficient rAAV vector for protein expression in cortical parvalbumin expressing interneurons
title_fullStr An efficient rAAV vector for protein expression in cortical parvalbumin expressing interneurons
title_full_unstemmed An efficient rAAV vector for protein expression in cortical parvalbumin expressing interneurons
title_short An efficient rAAV vector for protein expression in cortical parvalbumin expressing interneurons
title_sort efficient raav vector for protein expression in cortical parvalbumin expressing interneurons
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9596399/
https://www.ncbi.nlm.nih.gov/pubmed/36284123
http://dx.doi.org/10.1038/s41598-022-21867-0
work_keys_str_mv AT tkatchtatiana anefficientraavvectorforproteinexpressionincorticalparvalbuminexpressinginterneurons
AT rysevaitekyguolienekristina anefficientraavvectorforproteinexpressionincorticalparvalbuminexpressinginterneurons
AT sabeckisignas anefficientraavvectorforproteinexpressionincorticalparvalbuminexpressinginterneurons
AT sabeckienedeimante anefficientraavvectorforproteinexpressionincorticalparvalbuminexpressinginterneurons
AT pauzadainiush anefficientraavvectorforproteinexpressionincorticalparvalbuminexpressinginterneurons
AT baranauskasgytis anefficientraavvectorforproteinexpressionincorticalparvalbuminexpressinginterneurons
AT tkatchtatiana efficientraavvectorforproteinexpressionincorticalparvalbuminexpressinginterneurons
AT rysevaitekyguolienekristina efficientraavvectorforproteinexpressionincorticalparvalbuminexpressinginterneurons
AT sabeckisignas efficientraavvectorforproteinexpressionincorticalparvalbuminexpressinginterneurons
AT sabeckienedeimante efficientraavvectorforproteinexpressionincorticalparvalbuminexpressinginterneurons
AT pauzadainiush efficientraavvectorforproteinexpressionincorticalparvalbuminexpressinginterneurons
AT baranauskasgytis efficientraavvectorforproteinexpressionincorticalparvalbuminexpressinginterneurons