Biology subjects

Kietrys, A. M.

Publications and source records attributed to Kietrys, A. M..

4 recordsLinked to original sources

bioRxiv2026

Reversible RNA Acylating Reagents with Nitro Reduction Strategy

We developed a series of nitro reduction-reversible acylating reagents. Following optimization of the acylation conditions, these reagents were tested for deacylation with sodium dithionite in vitro. We applied this reversible acylation to modulate RNAzyme-mediated pre-tRNA maturation, demonstrating its ability to regulate RNA-RNA interactions. Furthermore, the in vitro reversible acylation of EGFP mRNA indicated effective control of its translational activity. To explore cellular applications, we validated NQO1-mediated deacylation in vitro and then induced hypoxia in HepG2 cells using cobalt chloride, thereby reactivating the function of acylated EGFP mRNA via endogenous NQO1. Overall, this study highlights the potential for developing nitro reduction-reversible acylation as a new strategy for RNA functional control and RNA-based drug modification.

biochemistry↗

bioRxiv2025

Unlocking efficiency: Native circular RNA surpass linear isoforms in RNase P activity

RNase P, one of the earliest enzymes identified with RNA as its catalytic component, has been extensively studied across all three domains of life. In this research, we unveil circular isoforms of RNase P RNA within bacterial, fungal, and human cell lines. Comparing the bacterial variant, circM1, with its linear counterpart under diverse conditions revealed its enhanced temperature resistance and superior tolerance to Mn2+. Moreover, our findings suggest distinct protein associations for both isoforms in the presence of FBS. The human counterpart, circH1, was proved to be active in cellulo.

cell biology↗

bioRxiv2024

RNA functional control by hydrolysis reversible acylation

Reversible 2'-OH acylation is a powerful strategy for switching RNA function, but existing systems often rely on nonphysiological or cytotoxic triggers for deacylation. Here we present EST1A, a hydrolysis-responsive 2'-OH acylating reagent whose RNA adducts are efficiently removed by endogenous esterases in vitro and in cellulo. EST1A acylates model oligonucleotides, an EGFP-targeting antisense strand, and reporter mRNAs, thereby modulating their activity; notably, the acylated antisense strand shows enhanced EGFP knockdown in HepG2 cells. By tuning carboxylesterase and cholinesterase activity and comparing EST1A-acylated mCherry mRNA across noncancerous and cancer-derived cell lines, we reveal a positive correlation between intracellular esterase activity and functional recovery of acylated RNA. These results establish EST1A-mediated, hydrolysis-responsive 2'-OH acylation as a simple platform for enzyme-guided, cell-selective activation of RNA function and point toward esterase-activated RNA therapeutics. Entry for the Table of Contents O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=118 SRC="FIGDIR/small/610419v2_ufig1.gif" ALT="Figure 1"> View larger version (14K): org.highwire.dtl.DTLVardef@1cbce3org.highwire.dtl.DTLVardef@b51bc4org.highwire.dtl.DTLVardef@31e96dorg.highwire.dtl.DTLVardef@d42578_HPS_FORMAT_FIGEXP M_FIG Liu et al. introduce EST1A, a hydrolysis-responsive 2'-OH acylating reagent whose RNA adducts are removed by endogenous esterases or histidine, enabling reversible control of RNA function. By exploiting differences in esterase activity between noncancerous and cancer-derived cell lines, EST1A-treated mRNA exhibits enzyme-guided and cell-selective translational reactivation. C_FIG

cancer biology↗

bioRxiv2022

Pervasive Transcriptome Interactions of Protein-Targeted Drugs

The off-target toxicity of drugs targeted to proteins imparts substantial health and economic costs. Proteome interaction studies can reveal off-target effects with unintended proteins; however, little attention has been paid to intracellular RNAs as potential off targets that may contribute to toxicity. To begin to assess this, we developed a reactivity-based RNA profiling (RBRP) methodology, and applied it to uncover transcriptome interactions of a set of FDA-approved small-molecule drugs in vivo. We show that these protein-targeted drugs pervasively interact with the human transcriptome and can exert unintended biological effects on RNA function. In addition, we show that many off-target interactions occur at RNA loci associated with protein binding and structural changes, allowing us to generate hypotheses to infer the biological consequences of RNA off-target binding. The results suggest that rigorous characterization of drugs' transcriptome interactions may help assess target specificity and potentially avoid toxicity and clinical failures.

pharmacology and toxicology↗