BFF-piRNA-seq reveals conserved processing hotspots in flies and mice
Abstract
PIWI-interacting RNAs (piRNAs) protect germline genome integrity and fertility, but quantitative profiling of mature PIWI-bound RNAs remains technically challenging. Here we develop BFF-piRNA-seq, which ligates sequencing adapters directly to RNAs on immobilized PIWI complexes, eliminating preparative polyacrylamide gel electrophoresis and associated size-selection bias. Split-sample comparisons of on- and off-PIWI ligation show highly concordant length distributions and no detectable terminal sequence bias, and the method reproducibly profiles distinct PIWI-piRNA complexes in mouse and Drosophila . Eliminating gel selection recovers a broader population of Drosophila Piwi-piRNAs than conventional workflows. Across flies and mice, quantitative profiles reveal a conserved landscape across piRNA clusters in which narrow hotspots generate abundant families of sequence-related piRNAs with 5′ microheterogeneity. Together, these data establish BFF-piRNA-seq as an accessible, quantitative approach for profiling bona fide piRNAs and uncover conserved principles governing piRNA selection during biogenesis and maturation.
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BibTeX
@misc{haase2026bffpirnaseq,
title = {{BFF-piRNA-seq reveals conserved processing hotspots in flies and mice}},
author = {Astrid Haase and Markus Hafner and Parthena Konstantinidou and Franziska Ahrend and Brian Galletta and Angel Jaimes and Dimitrios Anastasakis and Nasser Rusan},
howpublished = {Research Square},
year = {2026},
month = oct,
doi = {10.21203/rs.3.rs-11307628/v1},
url = {https://doi.org/10.21203/rs.3.rs-11307628/v1},
note = {CC BY 4.0},
}
RIS
TY - UNPB
TI - BFF-piRNA-seq reveals conserved processing hotspots in flies and mice
AU - Astrid Haase
AU - Markus Hafner
AU - Parthena Konstantinidou
AU - Franziska Ahrend
AU - Brian Galletta
AU - Angel Jaimes
AU - Dimitrios Anastasakis
AU - Nasser Rusan
PY - 2026
DA - 2026/10/26
PB - Research Square
DO - 10.21203/rs.3.rs-11307628/v1
UR - https://doi.org/10.21203/rs.3.rs-11307628/v1
AB - PIWI-interacting RNAs (piRNAs) protect germline genome integrity and fertility, but quantitative profiling of mature PIWI-bound RNAs remains technically challenging. Here we develop BFF-piRNA-seq, which ligates sequencing adapters directly to RNAs on immobilized PIWI complexes, eliminating preparative polyacrylamide gel electrophoresis and associated size-selection bias. Split-sample comparisons of on- and off-PIWI ligation show highly concordant length distributions and no detectable terminal sequence bias, and the method reproducibly profiles distinct PIWI-piRNA complexes in mouse and Drosophila . Eliminating gel selection recovers a broader population of Drosophila Piwi-piRNAs than conventional workflows. Across flies and mice, quantitative profiles reveal a conserved landscape across piRNA clusters in which narrow hotspots generate abundant families of sequence-related piRNAs with 5′ microheterogeneity. Together, these data establish BFF-piRNA-seq as an accessible, quantitative approach for profiling bona fide piRNAs and uncover conserved principles governing piRNA selection during biogenesis and maturation.
SN - 2693-5015
ER -