Helper-dependent adenovirus-mediated short hairpin RNA expression in the liver activates the interferon response

Scott R. Witting, Matthew Brown, Romil Saxena, Sarah Nabinger, Nuria Morral

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The use of RNA interference has proven to be an effective means to study the function of genes. Constitutive synthesis of small interfering RNA molecules can be accomplished with the use of viral vectors expressing short hairpin RNA (shRNA). Binding of shRNA to the target mRNA promotes transcript degradation. So far, little is known about the effects that shRNA induce in vivo. To determine the feasibility of using helper-dependent adenoviral vectors for expression of shRNA in liver, we have designed an shRNA construct to mouse fabp5 (fatty acid-binding protein 5). Intravenous administration of this vector resulted in ∼75% silencing of fabp5. Increasing the dose of vector did not result in higher levels of silencing, indicating that there is a threshold for the level of knockdown that can be achieved. Synthesis of high levels of shRNA molecules did not alter the levels of cellular micro-RNA, such as miR-122 and let-7a, suggesting that the exportin-5 pathway was not affected. However, high level shRNA expression resulted in activation of the interferon response. Thus, an important consideration when using shRNA-based vectors in vivo is to closely monitor signs of interferon-stimulated gene expression, since a narrow window exists between gene silencing efficacy and nonspecific effects.

Original languageEnglish
Pages (from-to)2120-2128
Number of pages9
JournalJournal of Biological Chemistry
Volume283
Issue number4
DOIs
StatePublished - Jan 25 2008

Fingerprint

Adenoviridae
Liver
Interferons
Small Interfering RNA
Fatty Acid-Binding Proteins
Genes
Karyopherins
Molecules
RNA Stability
Gene Silencing
RNA Interference
MicroRNAs
Gene expression
Intravenous Administration
Chemical activation
RNA
Gene Expression
Degradation
Messenger RNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Helper-dependent adenovirus-mediated short hairpin RNA expression in the liver activates the interferon response. / Witting, Scott R.; Brown, Matthew; Saxena, Romil; Nabinger, Sarah; Morral, Nuria.

In: Journal of Biological Chemistry, Vol. 283, No. 4, 25.01.2008, p. 2120-2128.

Research output: Contribution to journalArticle

@article{c123d6eecdf644789e7631c3be071fb3,
title = "Helper-dependent adenovirus-mediated short hairpin RNA expression in the liver activates the interferon response",
abstract = "The use of RNA interference has proven to be an effective means to study the function of genes. Constitutive synthesis of small interfering RNA molecules can be accomplished with the use of viral vectors expressing short hairpin RNA (shRNA). Binding of shRNA to the target mRNA promotes transcript degradation. So far, little is known about the effects that shRNA induce in vivo. To determine the feasibility of using helper-dependent adenoviral vectors for expression of shRNA in liver, we have designed an shRNA construct to mouse fabp5 (fatty acid-binding protein 5). Intravenous administration of this vector resulted in ∼75{\%} silencing of fabp5. Increasing the dose of vector did not result in higher levels of silencing, indicating that there is a threshold for the level of knockdown that can be achieved. Synthesis of high levels of shRNA molecules did not alter the levels of cellular micro-RNA, such as miR-122 and let-7a, suggesting that the exportin-5 pathway was not affected. However, high level shRNA expression resulted in activation of the interferon response. Thus, an important consideration when using shRNA-based vectors in vivo is to closely monitor signs of interferon-stimulated gene expression, since a narrow window exists between gene silencing efficacy and nonspecific effects.",
author = "Witting, {Scott R.} and Matthew Brown and Romil Saxena and Sarah Nabinger and Nuria Morral",
year = "2008",
month = "1",
day = "25",
doi = "10.1074/jbc.M704178200",
language = "English",
volume = "283",
pages = "2120--2128",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "4",

}

TY - JOUR

T1 - Helper-dependent adenovirus-mediated short hairpin RNA expression in the liver activates the interferon response

AU - Witting, Scott R.

AU - Brown, Matthew

AU - Saxena, Romil

AU - Nabinger, Sarah

AU - Morral, Nuria

PY - 2008/1/25

Y1 - 2008/1/25

N2 - The use of RNA interference has proven to be an effective means to study the function of genes. Constitutive synthesis of small interfering RNA molecules can be accomplished with the use of viral vectors expressing short hairpin RNA (shRNA). Binding of shRNA to the target mRNA promotes transcript degradation. So far, little is known about the effects that shRNA induce in vivo. To determine the feasibility of using helper-dependent adenoviral vectors for expression of shRNA in liver, we have designed an shRNA construct to mouse fabp5 (fatty acid-binding protein 5). Intravenous administration of this vector resulted in ∼75% silencing of fabp5. Increasing the dose of vector did not result in higher levels of silencing, indicating that there is a threshold for the level of knockdown that can be achieved. Synthesis of high levels of shRNA molecules did not alter the levels of cellular micro-RNA, such as miR-122 and let-7a, suggesting that the exportin-5 pathway was not affected. However, high level shRNA expression resulted in activation of the interferon response. Thus, an important consideration when using shRNA-based vectors in vivo is to closely monitor signs of interferon-stimulated gene expression, since a narrow window exists between gene silencing efficacy and nonspecific effects.

AB - The use of RNA interference has proven to be an effective means to study the function of genes. Constitutive synthesis of small interfering RNA molecules can be accomplished with the use of viral vectors expressing short hairpin RNA (shRNA). Binding of shRNA to the target mRNA promotes transcript degradation. So far, little is known about the effects that shRNA induce in vivo. To determine the feasibility of using helper-dependent adenoviral vectors for expression of shRNA in liver, we have designed an shRNA construct to mouse fabp5 (fatty acid-binding protein 5). Intravenous administration of this vector resulted in ∼75% silencing of fabp5. Increasing the dose of vector did not result in higher levels of silencing, indicating that there is a threshold for the level of knockdown that can be achieved. Synthesis of high levels of shRNA molecules did not alter the levels of cellular micro-RNA, such as miR-122 and let-7a, suggesting that the exportin-5 pathway was not affected. However, high level shRNA expression resulted in activation of the interferon response. Thus, an important consideration when using shRNA-based vectors in vivo is to closely monitor signs of interferon-stimulated gene expression, since a narrow window exists between gene silencing efficacy and nonspecific effects.

UR - http://www.scopus.com/inward/record.url?scp=38349149279&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=38349149279&partnerID=8YFLogxK

U2 - 10.1074/jbc.M704178200

DO - 10.1074/jbc.M704178200

M3 - Article

C2 - 18025090

AN - SCOPUS:38349149279

VL - 283

SP - 2120

EP - 2128

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 4

ER -