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Induction of RNAi Responses by Short Left-Handed Hairpin RNAi Triggers.

ABSTRACT: Small double-stranded, left-handed hairpin (LHP) RNAs containing a 5'-guide-loop-passenger-3' structure induce RNAi responses by a poorly understood mechanism. To explore LHPs, we synthesized fully 2'-modified LHP RNAs targeting multiple genes and found all to induce robust RNAi responses. Deletion of the loop and nucleotides at the 5'-end of the equivalent passenger strand resulted in a smaller LHP that still induced strong RNAi responses. Surprisingly, progressive deletion of up to 10 nucleotides from the 3'-end of the guide strand resulted in a 32mer LHP capable of inducing robust RNAi responses. However, further guide strand deletion inhibited LHP activity, thereby defining the minimal length guide targeting length to 13 nucleotides. To dissect LHP processing, we examined LHP species that coimmunoprecipitated with Argonaute 2 (Ago2), the catalytic core of RNA-induced silencing complex, and found that the Ago2-associated processed LHP species was of a length that correlated with Ago2 cleavage of the passenger strand. Placement of a blocking 2'-OMe blocking modification at the LHP predicted Ago2 cleavage site resulted in an intact LHP loaded into Ago2 and no RNAi response. Taken together, these data argue that in the absence of a substantial loop, this novel class of small LHP RNAs enters the RNAi pathway by a Dicer-independent mechanism that involves Ago2 cleavage and results in an extended guide strand. This work establishes LHPs as an alternative RNAi trigger that can be produced from a single synthesis for potential use as an RNAi therapeutic.

SUBMITTER: Hagopian JC

PROVIDER: S-EPMC5649124 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature

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Induction of RNAi Responses by Short Left-Handed Hairpin RNAi Triggers.

Hagopian Jonathan C JC Hamil Alexander S AS van den Berg Arjen A Meade Bryan R BR Eguchi Akiko A Palm-Apergi Caroline C Dowdy Steven F SF

Nucleic acid therapeutics 20170921 5

Small double-stranded, left-handed hairpin (LHP) RNAs containing a 5'-guide-loop-passenger-3' structure induce RNAi responses by a poorly understood mechanism. To explore LHPs, we synthesized fully 2'-modified LHP RNAs targeting multiple genes and found all to induce robust RNAi responses. Deletion of the loop and nucleotides at the 5'-end of the equivalent passenger strand resulted in a smaller LHP that still induced strong RNAi responses. Surprisingly, progressive deletion of up to 10 nucleoti ...[more]

PMID: 28933656

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Project description:The purpose of this study was to examine performance advantages associated with batting stance, in the form of left- vs. right-handed dominant stance, and orthodox vs. reverse stance, of talented junior cricket batters within age-restricted competitions. Data were sourced from the national male younger age competition (YAC; Under-17; n = 237) and older age competition (OAC; Under-19; n = 302), as well as female YAC (Under-15; n = 234) and OAC (Under-18; n = 260) over a 4-year period. Left-hand dominant (LHD) batters were consistently overrepresented in the male YAC (Right: 69.2%; Left: 30.8%) and OAC (Right: 68.2%; Left: 31.8%) compared with the expected general population distribution. Male LHD batters exhibited a significantly (p < 0.05) higher batting aggregate (YAC: 116.82 ± 84.75 vs. 137.84 ± 89.74; OAC: 117.07 ± 89.00 vs. 146.28 ± 95.99), scored more runs (YAC: 19.65 ± 12.32 vs. 23.96 ± 14.71; OAC: 19.27 ± 12.61 vs. 23.98 ± 14.15), spent more time batting (YAC: 45.33 ± 25.89 min vs. 54.59 ± 28.62 min; OAC: 39.80 ± 21.79 min vs. 49.33 ± 27.41 min), and scored more boundary-4s per game (YAC: 1.83 ± 1.40 vs. 2.44 ± 1.87; OAC: 1.76 ± 1.32 vs. 2.19 ± 1.83), across both YAC and OAC groups with small effect sizes. No overrepresentation was present for either female group (YAC, Right: 88.5%/Left: 11.5%; OAC, Right: 90.0%/Left: 10.0%). Female LHD batters exhibited significantly higher batting aggregate (68.97 ± 53.17 vs. 102.96 ± 73.48), batting average (13.24 ± 10.88 vs. 17.75 ± 12.28), and spent more time batting per game (25.52 ± 15.08 vs. 37.75 ± 26.76 min), but only at the OAC level with small-moderate effects sizes. Finally, there were few performance advantages identified to batting with a reverse stance, with further work needed to clarify any potential biomechanical benefits. Team selection practices may exploit the left-handed advantage by over-selecting talented left-handed junior cricketers. Practical implications for coaches include creating practice environments that negate the negative frequency-dependent selection, such as providing more practice opportunities for their players against left-handed opponents.

Dataset Information

Induction of RNAi Responses by Short Left-Handed Hairpin RNAi Triggers.

Publications

Induction of RNAi Responses by Short Left-Handed Hairpin RNAi Triggers.

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