Iwasaki, S. et al. Hsc70/Hsp90 chaperone machinery mediates ATP-dependent RISC loading of small RNA duplexes. Mol. Cell 39, 292–299 (2010).
Miyoshi, T., Takeuchi, A., Siomi, H. & Siomi, M. C. A direct role for Hsp90 in pre-RISC formation in Drosophila. Nat. Struct. Mol. Biol. 17, 1024–1026 (2010).
Iki, T. et al. In vitro assembly of plant RNA-induced silencing complexes facilitated by molecular chaperone HSP90. Mol. Cell 39, 282–291 (2010).
Kim, H., Lee, Y.-Y. & Kim, V. N. The biogenesis and regulation of animal microRNAs. Nat. Rev. Mol. Cell Biol. 26, 276–296 (2025).
Matranga, C., Tomari, Y., Shin, C., Bartel, D. P. & Zamore, P. D. Passenger-strand cleavage facilitates assembly of siRNA into Ago2-containing RNAi enzyme complexes. Cell 123, 607–620 (2005).
Schirle, N. T. & MacRae, I. J. The crystal structure of human Argonaute2. Science 336, 1037–1040 (2012).
Elkayam, E. et al. The structure of human argonaute-2 in complex with miR-20a. Cell 150, 233 (2012).
Song, J.-J., Smith, S. K., Hannon, G. J. & Joshua-Tor, L. Crystal structure of Argonaute and its implications for RISC slicer activity. Science 305, 1434–1437 (2004).
Iwakawa, H.-O. & Tomari, Y. Life of RISC: formation, action, and degradation of RNA-induced silencing complex. Mol. Cell 82, 30–43 (2022).
Kirschke, E., Goswami, D., Southworth, D., Griffin, P. R. & Agard, D. A. Glucocorticoid receptor function regulated by coordinated action of the Hsp90 and Hsp70 chaperone cycles. Cell 157, 1685–1697 (2014).
Wang, R. Y.-R. et al. Structure of Hsp90-Hsp70-Hop-GR reveals the Hsp90 client-loading mechanism. Nature 601, 460–464 (2022).
Noddings, C. M., Wang, R. Y.-R., Johnson, J. L. & Agard, D. A. Structure of Hsp90-p23-GR reveals the Hsp90 client-remodelling mechanism. Nature 601, 465–469 (2022).
Iwasaki, S. et al. Defining fundamental steps in the assembly of the Drosophila RNAi enzyme complex. Nature 521, 533–536 (2015).
Naruse, K., Matsuura-Suzuki, E., Watanabe, M., Iwasaki, S. & Tomari, Y. In vitro reconstitution of chaperone-mediated human RISC assembly. RNA 24, 6–11 (2018).
Tsuboyama, K., Tadakuma, H. & Tomari, Y. Conformational activation of Argonaute by distinct yet coordinated actions of the Hsp70 and Hsp90 chaperone systems. Mol. Cell 70, 722–729 (2018).
Liu, J. et al. Argonaute2 is the catalytic engine of mammalian RNAi. Science 305, 1437–1441 (2004).
Landthaler, M. et al. Molecular characterization of human Argonaute-containing ribonucleoprotein complexes and their bound target mRNAs. RNA 14, 2580–2596 (2008).
Verba, K. A. et al. Atomic structure of Hsp90-Cdc37-Cdk4 reveals that Hsp90 traps and stabilizes an unfolded kinase. Science 352, 1542–1547 (2016).
Suzuki, H. I. et al. Small-RNA asymmetry is directly driven by mammalian Argonautes. Nat. Struct. Mol. Biol. 22, 512–521 (2015).
Winter, J. & Diederichs, S. Argonaute-3 activates the let-7a passenger strand microRNA. RNA Biol. 10, 1631–1643 (2013).
Kobayashi, H., Shoji, K., Kiyokawa, K., Negishi, L. & Tomari, Y. Iruka eliminates dysfunctional Argonaute by selective ubiquitination of its empty state. Mol. Cell 73, 119–129 (2019).
Nakanishi, K., Weinberg, D. E., Bartel, D. P. & Patel, D. J. Structure of yeast Argonaute with guide RNA. Nature 486, 368–374 (2012).
Lee, K. et al. The structure of an Hsp90-immunophilin complex reveals cochaperone recognition of the client maturation state. Mol. Cell 81, 3496–3508 (2021).
Gruszczyk, J. et al. Cryo-EM structure of the agonist-bound Hsp90-XAP2-AHR cytosolic complex. Nat. Commun. 13, 7010 (2022).
García-Alonso, S. et al. Structure of the RAF1-HSP90-CDC37 complex reveals the basis of RAF1 regulation. Mol. Cell 82, 3438–3452 (2022).
Oberoi, J. et al. HSP90-CDC37-PP5 forms a structural platform for kinase dephosphorylation. Nat. Commun. 13, 7343 (2022).
Schirle, N. T., Sheu-Gruttadauria, J., Chandradoss, S. D., Joo, C. & MacRae, I. J. Water-mediated recognition of t1-adenosine anchors Argonaute2 to microRNA targets. eLife 4, e07646 (2015).
Schirle, N. T., Sheu-Gruttadauria, J. & MacRae, I. J. Structural basis for microRNA targeting. Science 346, 608–613 (2014).
Mohamed, A. A., Wang, P. Y., Bartel, D. P. & Vos, S. M. The structural basis for RNA slicing by human Argonaute2. Cell Rep. 44, 115166 (2025).
Rinaldi, S., Colombo, G. & Paladino, A. Mechanistic model for the Hsp90-driven opening of human Argonaute. J. Chem. Inf. Model. 60, 1469–1480 (2020).
Abramson, J. et al. Accurate structure prediction of biomolecular interactions with AlphaFold 3. Nature 630, 493–500 (2024).
Sheu-Gruttadauria, J. & MacRae, I. J. Phase transitions in the assembly and function of human miRISC. Cell 173, 946–957 (2018).
Noddings, C. M., Johnson, J. L. & Agard, D. A. Cryo-EM reveals how Hsp90 and FKBP immunophilins co-regulate the glucocorticoid receptor. Nat. Struct. Mol. Biol. 30, 1867–1877 (2023).
Jaime-Garza, M. et al. Hsp90 provides a platform for kinase dephosphorylation by PP5. Nat. Commun. 14, 2197 (2023).
Ali, M. M. U. et al. Crystal structure of an Hsp90-nucleotide-p23/Sba1 closed chaperone complex. Nature 440, 1013–1017 (2006).
Taipale, M. et al. Quantitative analysis of HSP90-client interactions reveals principles of substrate recognition. Cell 150, 987–1001 (2012).
Martinez, J., Patkaniowska, A., Urlaub, H., Lührmann, R. & Tuschl, T. Single-stranded antisense siRNAs guide target RNA cleavage in RNAi. Cell 110, 563–574 (2002).
Schirle, N. T. et al. Structural analysis of human Argonaute-2 bound to a modified siRNA guide. J. Am. Chem. Soc. 138, 8694–8697 (2016).
Tang, Q. & Khvorova, A. RNAi-based drug design: considerations and future directions. Nat. Rev. Drug Discov. 23, 341–364 (2024).
Foster, D. J. et al. Advanced siRNA designs further improve in vivo performance of GalNAc-siRNA conjugates. Mol. Ther. 26, 708–717 (2018).
Zheng, J. et al. Single modification at position 14 of siRNA strand abolishes its gene-silencing activity by decreasing both RISC loading and target degradation. FASEB J. 27, 4017–4026 (2013).
Hu, B. et al. Therapeutic siRNA: state of the art. Signal Transduct. Target. Ther. 5, 101 (2020).
Rosenzweig, R., Nillegoda, N. B., Mayer, M. P. & Bukau, B. The Hsp70 chaperone network. Nat. Rev. Mol. Cell Biol. 20, 665–680 (2019).
Kwak, P. B. & Tomari, Y. The N domain of Argonaute drives duplex unwinding during RISC assembly. Nat. Struct. Mol. Biol. 19, 145–151 (2012).
Pare, J. M., LaPointe, P. & Hobman, T. C. Hsp90 cochaperones p23 and FKBP4 physically interact with hAgo2 and activate RNA interference-mediated silencing in mammalian cells. Mol. Biol. Cell 24, 2303–2310 (2013).
Martinez, N. J., Chang, H.-M., Borrajo, J., de, R. & Gregory, R. I. The co-chaperones Fkbp4/5 control Argonaute2 expression and facilitate RISC assembly. RNA 19, 1583–1593 (2013).
Yamaguchi, S. et al. Structure of the Dicer-2–R2D2 heterodimer bound to a small RNA duplex. Nature 607, 393–398 (2022).
Broto, P., Moreau, G. & Vandycke, C. Molecular structures: perception, autocorrelation descriptor and sar studies. Perception of molecules: topological structure and 3-dimensional structure. European J. Med. Chem. 19, 61–65 (1984).
Prakash, T. P. et al. Targeted delivery of antisense oligonucleotides to hepatocytes using triantennary N-acetyl galactosamine improves potency 10-fold in mice. Nucleic Acids Res. 42, 8796–8807 (2014).
Csermely, P. et al. ATP induces a conformational change of the 90-kDa heat shock protein (hsp90). J. Biol. Chem. 268, 1901–1907 (1993).
Punjani, A., Rubinstein, J. L., Fleet, D. J. & Brubaker, M. A. cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination. Nat. Methods 14, 290–296 (2017).
Goddard, T. D. et al. UCSF ChimeraX: meeting modern challenges in visualization and analysis. Protein Sci. 27, 14–25 (2018).
Srivastava, D., Yadav, R. P., Singh, S., Boyd, K. & Artemyev, N. O. Unique interface and dynamics of the complex of HSP90 with a specialized cochaperone AIPL1. Structure 31, 309–317 (2023).
Lee, Y.-Y., Lee, H., Kim, H., Kim, V. N. & Roh, S.-H. Structure of the human DICER-pre-miRNA complex in a dicing state. Nature 615, 331–338 (2023).
Emsley, P. & Cowtan, K. Coot: model-building tools for molecular graphics. Acta Crystallogr. D Biol. Crystallogr. 60, 2126–2132 (2004).
Liebschner, D. et al. Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix. Acta Crystallogr. D 75, 861–877 (2019).

