Deep Sequencing Analyses of DsiRNAs Reveal the Influence of 3′ Terminal Overhangs on Dicing Polarity, Strand Selectivity, and RNA Editing of siRNAs
Identifieur interne : 001301 ( Pmc/Checkpoint ); précédent : 001300; suivant : 001302Deep Sequencing Analyses of DsiRNAs Reveal the Influence of 3′ Terminal Overhangs on Dicing Polarity, Strand Selectivity, and RNA Editing of siRNAs
Auteurs : Jiehua Zhou [États-Unis] ; Min-Sun Song [États-Unis] ; Ashley M. Jacobi [États-Unis] ; Mark A. Behlke [États-Unis] ; Xiwei Wu [États-Unis] ; John J. Rossi [États-Unis]Source :
- Molecular therapy. Nucleic acids [ 2162-2531 ] ; 2012.
Abstract
25/27 Base duplex RNAs that are substrates for Dicer have been demonstrated to enhance
RNA interference (RNAi) potency and efficacy. Since the target sites are not always
equally susceptible to suppression by small interfering RNA (siRNA), not all 27-mer
duplexes that are processed into the corresponding conventional siRNAs show increased
potency. Thus random designing of Dicer-substrate siRNAs (DsiRNAs) may generate siRNAs
with poor RNAi due to unpredictable Dicer processing. Previous studies have demonstrated
that the 3
Url:
DOI: 10.1038/mtna.2012.6
PubMed: 23343928
PubMed Central: 3384246
Affiliations:
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on Dicing Polarity, Strand Selectivity, and RNA Editing of siRNAs</title>
<author><name sortKey="Zhou, Jiehua" sort="Zhou, Jiehua" uniqKey="Zhou J" first="Jiehua" last="Zhou">Jiehua Zhou</name>
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, Duarte, California,<country>USA</country>
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<author><name sortKey="Song, Min Sun" sort="Song, Min Sun" uniqKey="Song M" first="Min-Sun" last="Song">Min-Sun Song</name>
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, Duarte, California,<country>USA</country>
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<author><name sortKey="Jacobi, Ashley M" sort="Jacobi, Ashley M" uniqKey="Jacobi A" first="Ashley M" last="Jacobi">Ashley M. Jacobi</name>
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<author><name sortKey="Behlke, Mark A" sort="Behlke, Mark A" uniqKey="Behlke M" first="Mark A" last="Behlke">Mark A. Behlke</name>
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<author><name sortKey="Wu, Xiwei" sort="Wu, Xiwei" uniqKey="Wu X" first="Xiwei" last="Wu">Xiwei Wu</name>
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, Duarte, California,<country>USA</country>
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<author><name sortKey="Rossi, John J" sort="Rossi, John J" uniqKey="Rossi J" first="John J" last="Rossi">John J. Rossi</name>
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, Duarte, California,<country>USA</country>
</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Irell and Manella Graduate School of Biological Sciences, Beckman Research Institute of City of Hope</institution>
, Duarte, California,<country>USA</country>
</nlm:aff>
<country xml:lang="fr">États-Unis</country>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Deep Sequencing Analyses of DsiRNAs Reveal the Influence of 3′ Terminal Overhangs
on Dicing Polarity, Strand Selectivity, and RNA Editing of siRNAs</title>
<author><name sortKey="Zhou, Jiehua" sort="Zhou, Jiehua" uniqKey="Zhou J" first="Jiehua" last="Zhou">Jiehua Zhou</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope</institution>
, Duarte, California,<country>USA</country>
</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Song, Min Sun" sort="Song, Min Sun" uniqKey="Song M" first="Min-Sun" last="Song">Min-Sun Song</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope</institution>
, Duarte, California,<country>USA</country>
</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Jacobi, Ashley M" sort="Jacobi, Ashley M" uniqKey="Jacobi A" first="Ashley M" last="Jacobi">Ashley M. Jacobi</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Integrated DNA Technologies, Inc.</institution>
Coralville, Iowa,<country>USA</country>
</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Behlke, Mark A" sort="Behlke, Mark A" uniqKey="Behlke M" first="Mark A" last="Behlke">Mark A. Behlke</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Integrated DNA Technologies, Inc.</institution>
Coralville, Iowa,<country>USA</country>
</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Wu, Xiwei" sort="Wu, Xiwei" uniqKey="Wu X" first="Xiwei" last="Wu">Xiwei Wu</name>
<affiliation wicri:level="1"><nlm:aff id="aff3"><institution>Department of Molecular Medicine, Beckman Research Institute of City of Hope</institution>
, Duarte, California,<country>USA</country>
</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Rossi, John J" sort="Rossi, John J" uniqKey="Rossi J" first="John J" last="Rossi">John J. Rossi</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope</institution>
, Duarte, California,<country>USA</country>
</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Irell and Manella Graduate School of Biological Sciences, Beckman Research Institute of City of Hope</institution>
, Duarte, California,<country>USA</country>
</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
</analytic>
<series><title level="j">Molecular therapy. Nucleic acids</title>
<idno type="eISSN">2162-2531</idno>
<imprint><date when="2012">2012</date>
</imprint>
</series>
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<front><div type="abstract" xml:lang="en"><p>25/27 Base duplex RNAs that are substrates for Dicer have been demonstrated to enhance
RNA interference (RNAi) potency and efficacy. Since the target sites are not always
equally susceptible to suppression by small interfering RNA (siRNA), not all 27-mer
duplexes that are processed into the corresponding conventional siRNAs show increased
potency. Thus random designing of Dicer-substrate siRNAs (DsiRNAs) may generate siRNAs
with poor RNAi due to unpredictable Dicer processing. Previous studies have demonstrated
that the 3<bold>′</bold>
-overhang affects dicing cleavage site and the orientation of
Dicer entry. Moreover, an asymmetric 27-mer duplex having a 3<bold>′</bold>
two-nucleotide
overhang and 3<bold>′</bold>
-DNA residues on the blunt end has been rationally designed to
obtain greater efficacy. This asymmetric structure directs dicing to predictably yield a
single primary cleavage product. In the present study, we analyzed the <italic>in vitro</italic>
and intracellular dicing patterns of chemically synthesized duplex RNAs with different
3<bold>′</bold>
-overhangs. Consistent with previous studies, we observed that Dicer
preferentially processes these RNAs at a site 21–22 nucleotide (nt) from the
two-base 3<bold>′</bold>
-overhangs. We also observed that the direction and ability of
human Dicer to generate siRNAs can be partially or completely blocked by DNA residues at
the 3<bold>′</bold>
-termimi. To examine the effects of various 3<bold>′</bold>
-end
modifications on Dicer processing in cells, we employed Illumina Deep sequencing analyses
to unravel the fates of the asymmetric 27-mer duplexes. To validate the strand selection
process and knockdown capabilities we also conducted dual-luciferase psiCHECK reporter
assays to monitor the RNAi potencies of both the “sense” (S) and
“antisense” (AS) strands derived from these DsiRNAs. Consistent with our
<italic>in vitro</italic>
Dicer assays, the asymmetric duplexes were predictably processed into
desired primary cleavage products of 21–22-mers in cells. We also observed the
trimming of the 3<bold>′</bold>
end, especially when DNA residues were incorporated into
the overhangs and this trimming ultimately influenced the Dicer-cleavage site and RNAi
potency. Moreover, the observation that the most efficacious strand was the most abundant
revealed that the relative frequencies of each “S” or “AS” strand
are highly correlated with the silencing activity and strand selectivity. Collectively,
our data demonstrate that even though the only differences between a family of DsiRNAs was
the 3<bold>′</bold>
two-nuclotide overhang, dicing polarity and strand selectivity are
distinct depending upon the sequence and chemical nature of this overhang. Thus, it is
possible to predictably control dicing polarity and strand selectivity <italic>via</italic>
simply
changing the 3<bold>′</bold>
-end overhangs without altering the original duplex sequence.
These optimal design features of 3<bold>′</bold>
-overhangs might provide a facile approach
for rationally designing highly potent 25/27-mer DsiRNAs.</p>
</div>
</front>
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<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Mol Ther Nucleic Acids</journal-id>
<journal-id journal-id-type="iso-abbrev">Mol Ther Nucleic Acids</journal-id>
<journal-title-group><journal-title>Molecular therapy. Nucleic acids</journal-title>
</journal-title-group>
<issn pub-type="epub">2162-2531</issn>
<publisher><publisher-name>Nature Publishing Group</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">23343928</article-id>
<article-id pub-id-type="pmc">3384246</article-id>
<article-id pub-id-type="pii">mtna20126</article-id>
<article-id pub-id-type="doi">10.1038/mtna.2012.6</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Original Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>Deep Sequencing Analyses of DsiRNAs Reveal the Influence of 3′ Terminal Overhangs
on Dicing Polarity, Strand Selectivity, and RNA Editing of siRNAs</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Zhou</surname>
<given-names>Jiehua</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Song</surname>
<given-names>Min-Sun</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Jacobi</surname>
<given-names>Ashley M</given-names>
</name>
<xref ref-type="aff" rid="aff2">2</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Behlke</surname>
<given-names>Mark A</given-names>
</name>
<xref ref-type="aff" rid="aff2">2</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wu</surname>
<given-names>Xiwei</given-names>
</name>
<xref ref-type="aff" rid="aff3">3</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Rossi</surname>
<given-names>John J</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="aff" rid="aff4">4</xref>
</contrib>
<aff id="aff1"><label>1</label>
<institution>Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope</institution>
, Duarte, California,<country>USA</country>
</aff>
<aff id="aff2"><label>2</label>
<institution>Integrated DNA Technologies, Inc.</institution>
Coralville, Iowa,<country>USA</country>
</aff>
<aff id="aff3"><label>3</label>
<institution>Department of Molecular Medicine, Beckman Research Institute of City of Hope</institution>
, Duarte, California,<country>USA</country>
</aff>
<aff id="aff4"><label>4</label>
<institution>Irell and Manella Graduate School of Biological Sciences, Beckman Research Institute of City of Hope</institution>
, Duarte, California,<country>USA</country>
</aff>
</contrib-group>
<author-notes><corresp id="caf1"><label>*</label>
Department of Molecular and Cellular Biology, Beckman Research
Institute of City of Hope, 1500 East Duarte Road, Duarte, California 91010, USA. E-mail
address: <email>jrossi@coh.org</email>
</corresp>
</author-notes>
<pub-date pub-type="ppub"><month>04</month>
<year>2012</year>
</pub-date>
<pub-date pub-type="epub"><day>03</day>
<month>04</month>
<year>2012</year>
</pub-date>
<pub-date pub-type="pmc-release"><day>1</day>
<month>4</month>
<year>2012</year>
</pub-date>
<volume>1</volume>
<issue>4</issue>
<fpage>e17</fpage>
<lpage></lpage>
<history><date date-type="received"><day>23</day>
<month>01</month>
<year>2012</year>
</date>
<date date-type="rev-recd"><day>18</day>
<month>02</month>
<year>2012</year>
</date>
<date date-type="accepted"><day>03</day>
<month>04</month>
<year>2012</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2012 American Society of Gene & Cell Therapy</copyright-statement>
<copyright-year>2012</copyright-year>
<copyright-holder>American Society of Gene & Cell Therapy</copyright-holder>
<license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by-nc-nd/3.0/"><pmc-comment>author-paid</pmc-comment>
<license-p><italic>Molecular Therapy-Nucleic Acids</italic>
is an open-access journal published
by Nature Publishing Group. This work is licensed under the Creative Commons
Attribution-NonCommercial-No Derivative Works 3.0 Unported License. To view a copy of this
license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/</license-p>
</license>
</permissions>
<abstract><p>25/27 Base duplex RNAs that are substrates for Dicer have been demonstrated to enhance
RNA interference (RNAi) potency and efficacy. Since the target sites are not always
equally susceptible to suppression by small interfering RNA (siRNA), not all 27-mer
duplexes that are processed into the corresponding conventional siRNAs show increased
potency. Thus random designing of Dicer-substrate siRNAs (DsiRNAs) may generate siRNAs
with poor RNAi due to unpredictable Dicer processing. Previous studies have demonstrated
that the 3<bold>′</bold>
-overhang affects dicing cleavage site and the orientation of
Dicer entry. Moreover, an asymmetric 27-mer duplex having a 3<bold>′</bold>
two-nucleotide
overhang and 3<bold>′</bold>
-DNA residues on the blunt end has been rationally designed to
obtain greater efficacy. This asymmetric structure directs dicing to predictably yield a
single primary cleavage product. In the present study, we analyzed the <italic>in vitro</italic>
and intracellular dicing patterns of chemically synthesized duplex RNAs with different
3<bold>′</bold>
-overhangs. Consistent with previous studies, we observed that Dicer
preferentially processes these RNAs at a site 21–22 nucleotide (nt) from the
two-base 3<bold>′</bold>
-overhangs. We also observed that the direction and ability of
human Dicer to generate siRNAs can be partially or completely blocked by DNA residues at
the 3<bold>′</bold>
-termimi. To examine the effects of various 3<bold>′</bold>
-end
modifications on Dicer processing in cells, we employed Illumina Deep sequencing analyses
to unravel the fates of the asymmetric 27-mer duplexes. To validate the strand selection
process and knockdown capabilities we also conducted dual-luciferase psiCHECK reporter
assays to monitor the RNAi potencies of both the “sense” (S) and
“antisense” (AS) strands derived from these DsiRNAs. Consistent with our
<italic>in vitro</italic>
Dicer assays, the asymmetric duplexes were predictably processed into
desired primary cleavage products of 21–22-mers in cells. We also observed the
trimming of the 3<bold>′</bold>
end, especially when DNA residues were incorporated into
the overhangs and this trimming ultimately influenced the Dicer-cleavage site and RNAi
potency. Moreover, the observation that the most efficacious strand was the most abundant
revealed that the relative frequencies of each “S” or “AS” strand
are highly correlated with the silencing activity and strand selectivity. Collectively,
our data demonstrate that even though the only differences between a family of DsiRNAs was
the 3<bold>′</bold>
two-nuclotide overhang, dicing polarity and strand selectivity are
distinct depending upon the sequence and chemical nature of this overhang. Thus, it is
possible to predictably control dicing polarity and strand selectivity <italic>via</italic>
simply
changing the 3<bold>′</bold>
-end overhangs without altering the original duplex sequence.
These optimal design features of 3<bold>′</bold>
-overhangs might provide a facile approach
for rationally designing highly potent 25/27-mer DsiRNAs.</p>
</abstract>
<kwd-group><kwd>RNA interference</kwd>
<kwd>Dicer-substrate siRNA (DsiRNA)</kwd>
<kwd>Deep sequencing</kwd>
<kwd>3′-overhang</kwd>
<kwd>Dicing polarity</kwd>
</kwd-group>
</article-meta>
</front>
<floats-group><fig id="fig1"><label>Figure 1</label>
<caption><p><bold><italic>In vitro</italic>
Dicer processing of (a) 5</bold>
′
<bold>P</bold>
<sup><bold>32</bold>
</sup>
<bold>-end labeled sense or antisense (b) duplexes.</bold>
Two
types of experimental approaches are displayed as method A and B, respectively. The
dicing patterns are named by the direction of Dicer entering the duplex as left to right
(L-R) and right to left (R-L).</p>
</caption>
<graphic xlink:href="mtna20126f1"></graphic>
</fig>
<fig id="fig2"><label>Figure 2</label>
<caption><p><bold><italic>In vitro</italic>
Dicer cleavage of 5</bold>
′ <bold>P</bold>
<sup><bold>32</bold>
</sup>
<bold>-end
labeled sense or antisense strands.</bold>
Group I duplexes. The cleavage fragments that
result from dicing L-R and R-L were visualized autoradiographically following denaturing
gel electrophoresis of the Dicer-cleavage products: (a) sense and (b) antisense. The two
experimental approaches (method A and B) are defined in the legend to Figure 1.</p>
</caption>
<graphic xlink:href="mtna20126f2"></graphic>
</fig>
<fig id="fig3"><label>Figure 3</label>
<caption><p><bold>Silencing of <italic>Homo sapiens</italic>
transportin 3 (<italic>TNPO3</italic>
) by group I duplex
Dicer-substrate small interfering RNAs (DsiRNAs)</bold>
. HEK 293 cells were transfected
with 50 or 10 nmol/l of the experimental anti-<italic>TNPO3</italic>
duplex DsiRNAs.
<italic>TNPO3</italic>
mRNA levels were detected by quantitative real-time reverse
transcription (qRT-PCR). The data are normalized with GAPDH mRNA levels and represent
the average of three replicate assays.</p>
</caption>
<graphic xlink:href="mtna20126f3"></graphic>
</fig>
<fig id="fig4"><label>Figure 4</label>
<caption><p><bold><italic>In vitro</italic>
Dicer cleavage of 5</bold>
′ <bold>P</bold>
<sup><bold>32</bold>
</sup>
<bold>-end
labeled sense or antisense strands</bold>
. Group II asymmetric duplex Dicer-substrate
small interfering RNAs (DsiRNAs) and RNA interference (RNAi) potency. The cleavage
fragments that result from dicing L-R and R-L were visualized by autoradiography of the
P<sup>32</sup>
5′ end-labeled strands: (<bold>a</bold>
) sense and (<bold>b</bold>
)
antisense. Two experimental approaches (method A and B) and the dicing pattern are
defined the legend to Figure 1. (<bold>c</bold>
) Silencing of <italic>Homo sapiens</italic>
transportin 3 (<italic>TNPO3</italic>
) triggered by group II duplex RNAs. HEK 293 cells were
transfected with 50 or 10 nmol/l of the experimental anti-<italic>TNPO3</italic>
duplex
DsiRNAs. TNPO3 mRNA level were detected by quantitative real-time reverse transcription
(qRT-PCR). The data were normalized with GAPDH mRNA and represent the average of three
replicate assays.</p>
</caption>
<graphic xlink:href="mtna20126f4"></graphic>
</fig>
<fig id="fig5"><label>Figure 5</label>
<caption><p><bold>Illumina Deep sequence analyses of asymmetric 27-mer <italic>Homo sapiens</italic>
transportin 3 (<italic>TNPO3</italic>
) Dicer-substrate siRNAs (group II).</bold>
HEK 293 cells
were transfected with 10 nmol/l of the asymmetric group II RNA duplexes. Forty
hours post-transfection the total RNAs were isolated and prepared for Illumina Deep
sequencing. The data collection and alignment are described in Materials and Methods
section. (<bold>a</bold>
) The total reads and abundance of sense and antisense strands from
each duplex. (<bold>b</bold>
) The strand distribution was calculated as the ratio of the
abundance of antisense to sense. The ratio of antisense (AS) to sense (S) is ranked by
the 3′ overhang GG > GC, AA > CC, UU ≫ tt. (<bold>c</bold>
) The dicing
pattern L-R and R-L are as previously described. The L-R pattern generates the desired
siRNA species for target knockdown. Total reads from the top 10 antisense strands and
the abundance of L-R cleavage products from the antisense strand. (<bold>d</bold>
) Two types
of RNA editing: trimming of the 3′ end and post-transcriptional addition of
nucleotides at the 3′ or 5′ ends.</p>
</caption>
<graphic xlink:href="mtna20126f5"></graphic>
</fig>
<fig id="fig6"><label>Figure 6</label>
<caption><p><bold>IC<sub>50</sub>
values and strand selectivity of asymmetric 27-mer <italic>Homo
sapiens</italic>
transportin 3 (<italic>TNPO3</italic>
) Dicer-substrate siRNAs (group II).</bold>
(<bold>a</bold>
) IC<sub>50</sub>
values of the asymmetric 27-mer duplexes were determined
using the psiCHECK assays as described Materials and Methods section. When the antisense
or sense was used as “guide” strand, the target knockdown efficiency is
listed as the IC<sub>50</sub>
. (<bold>b</bold>
) The strand selectivity was calculated as the
ratio of IC<sub>50</sub>
values of the sense to antisense targets.</p>
</caption>
<graphic xlink:href="mtna20126f6"></graphic>
</fig>
<fig id="fig7"><label>Figure 7</label>
<caption><p><bold>Illumina Deep sequence analyses of asymmetric 27-mer heterogeneous nuclear
ribonucleoprotein H (<italic>hnRNP H1</italic>
) Dicer-substrate small interfering RNA
(siRNAs).</bold>
The data collection and alignment were as described above. (<bold>a</bold>
)
Total reads of the top 10 sense and antisense strands from each duplex. (<bold>b</bold>
) The
strand distribution was calculated as the ratio of the abundance of antisense to sense.
Ratio of antisense (AS) to sense (S) is ranked by the 3′ overhang GG > AA in
site 324 Dicer-substrate small interfering RNAs (DsiRNAs) and GG > CC in site 325
DsiRNAs. (<bold>c</bold>
) The dicing pattern L-R and R-L are as previously described. The L-R
model generates the desired siRNA species for target knockdown. The total reads of the
top 10 antisense strands and the abundance of L-R cleavage products are presented.</p>
</caption>
<graphic xlink:href="mtna20126f7"></graphic>
</fig>
<table-wrap id="tbl1"><label>Table 1</label>
<caption><title>The 21-mer siRNAs and 27-mer Dicer-substrate siRNAs targeting <italic>TNPO3</italic>
and the
target sequences are listed</title>
</caption>
<graphic xlink:href="mtna20126t1"></graphic>
</table-wrap>
<table-wrap id="tbl2"><label>Table 2</label>
<caption><title>Illumina Deep sequence analyses and IC<sub>50</sub>
values of asymmetric 27-mer
<italic>TNPO3</italic>
Dicer-substrate siRNAs (group II)</title>
</caption>
<graphic xlink:href="mtna20126t2"></graphic>
</table-wrap>
<table-wrap id="tbl3"><label>Table 3</label>
<caption><title>Deep sequence analyses and IC<sub>50</sub>
values of the <italic>hnRNP H1</italic>
25/27-mer
Dicer-substrate siRNAs</title>
</caption>
<graphic xlink:href="mtna20126t3"></graphic>
</table-wrap>
</floats-group>
</pmc>
<affiliations><list><country><li>États-Unis</li>
</country>
</list>
<tree><country name="États-Unis"><noRegion><name sortKey="Zhou, Jiehua" sort="Zhou, Jiehua" uniqKey="Zhou J" first="Jiehua" last="Zhou">Jiehua Zhou</name>
</noRegion>
<name sortKey="Behlke, Mark A" sort="Behlke, Mark A" uniqKey="Behlke M" first="Mark A" last="Behlke">Mark A. Behlke</name>
<name sortKey="Jacobi, Ashley M" sort="Jacobi, Ashley M" uniqKey="Jacobi A" first="Ashley M" last="Jacobi">Ashley M. Jacobi</name>
<name sortKey="Rossi, John J" sort="Rossi, John J" uniqKey="Rossi J" first="John J" last="Rossi">John J. Rossi</name>
<name sortKey="Rossi, John J" sort="Rossi, John J" uniqKey="Rossi J" first="John J" last="Rossi">John J. Rossi</name>
<name sortKey="Song, Min Sun" sort="Song, Min Sun" uniqKey="Song M" first="Min-Sun" last="Song">Min-Sun Song</name>
<name sortKey="Wu, Xiwei" sort="Wu, Xiwei" uniqKey="Wu X" first="Xiwei" last="Wu">Xiwei Wu</name>
</country>
</tree>
</affiliations>
</record>
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