Oligonucleotide–Peptide Complexes: Phase Control by Hybridization
Abstract
When oppositely charged polymers are mixed, counterion release drives phase separation; understanding this process is a key unsolved problem in polymer science and biophysical chemistry, particularly for nucleic acids, polyanions whose biological functions are intimately related to their high charge density. In the cell, complexation by basic proteins condenses DNA into chromatin, and membraneless organelles formed by liquid-liquid phase separation of RNA and proteins perform vital functions and have been linked to disease. Electrostatic interactions are also the primary method used for assembly of nanoparticles to deliver therapeutic nucleic acids into cells. This paper describes complexation experiments with oligonucleotides and cationic peptides spanning a wide range of polymer lengths, concentrations, and structures, including RNA and methylphosphonate backbones. We find that the phase of the complexes is controlled by the hybridization state of the nucleic acid, with double-stranded nucleic acids forming solid precipitates while single-stranded oligonucleotides form liquid coacervates, apparently due to their lower charge density. Adding salt "melts" precipitates into coacervates, and oligonucleotides in coacervates remain competent for sequence-specific hybridization and phase change, suggesting the possibility of environmentally responsive complexes and nanoparticles for therapeutic or sensing applications.
- Authors:
-
[1];
[2];
- Univ. of Chicago, IL (United States). Inst. for Molecular Engineering
- Univ. of Chicago, IL (United States). Dept. of Chemistry
- Univ. of Central Florida, Orlando, FL (United States). Dept. of Materials Science and Engineering
- Univ. of Puerto Rico at Rio Piedras, San Juan, PR (United States). Dept. of Biological Sciences
- Univ. of Chicago, IL (United States). Inst. for Molecular Engineering; Argonne National Lab. (ANL), Argonne, IL (United States). Inst. for Molecular Engineering
- Publication Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22). Materials Sciences & Engineering Division
- OSTI Identifier:
- 1461292
- Grant/Contract Number:
- AC02-06CH11357
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of the American Chemical Society
- Additional Journal Information:
- Journal Volume: 140; Journal Issue: 5; Journal ID: ISSN 0002-7863
- Publisher:
- American Chemical Society (ACS)
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; 36 MATERIALS SCIENCE
Citation Formats
Vieregg, Jeffrey R., Lueckheide, Michael, Marciel, Amanda B., Leon, Lorraine, Bologna, Alex J., Rivera, Josean Reyes, and Tirrell, Matthew V. Oligonucleotide–Peptide Complexes: Phase Control by Hybridization. United States: N. p., 2018.
Web. doi:10.1021/jacs.7b03567.
Vieregg, Jeffrey R., Lueckheide, Michael, Marciel, Amanda B., Leon, Lorraine, Bologna, Alex J., Rivera, Josean Reyes, & Tirrell, Matthew V. Oligonucleotide–Peptide Complexes: Phase Control by Hybridization. United States. https://doi.org/10.1021/jacs.7b03567
Vieregg, Jeffrey R., Lueckheide, Michael, Marciel, Amanda B., Leon, Lorraine, Bologna, Alex J., Rivera, Josean Reyes, and Tirrell, Matthew V. Tue .
"Oligonucleotide–Peptide Complexes: Phase Control by Hybridization". United States. https://doi.org/10.1021/jacs.7b03567. https://www.osti.gov/servlets/purl/1461292.
@article{osti_1461292,
title = {Oligonucleotide–Peptide Complexes: Phase Control by Hybridization},
author = {Vieregg, Jeffrey R. and Lueckheide, Michael and Marciel, Amanda B. and Leon, Lorraine and Bologna, Alex J. and Rivera, Josean Reyes and Tirrell, Matthew V.},
abstractNote = {When oppositely charged polymers are mixed, counterion release drives phase separation; understanding this process is a key unsolved problem in polymer science and biophysical chemistry, particularly for nucleic acids, polyanions whose biological functions are intimately related to their high charge density. In the cell, complexation by basic proteins condenses DNA into chromatin, and membraneless organelles formed by liquid-liquid phase separation of RNA and proteins perform vital functions and have been linked to disease. Electrostatic interactions are also the primary method used for assembly of nanoparticles to deliver therapeutic nucleic acids into cells. This paper describes complexation experiments with oligonucleotides and cationic peptides spanning a wide range of polymer lengths, concentrations, and structures, including RNA and methylphosphonate backbones. We find that the phase of the complexes is controlled by the hybridization state of the nucleic acid, with double-stranded nucleic acids forming solid precipitates while single-stranded oligonucleotides form liquid coacervates, apparently due to their lower charge density. Adding salt "melts" precipitates into coacervates, and oligonucleotides in coacervates remain competent for sequence-specific hybridization and phase change, suggesting the possibility of environmentally responsive complexes and nanoparticles for therapeutic or sensing applications.},
doi = {10.1021/jacs.7b03567},
journal = {Journal of the American Chemical Society},
number = 5,
volume = 140,
place = {United States},
year = {Tue Jan 09 00:00:00 EST 2018},
month = {Tue Jan 09 00:00:00 EST 2018}
}
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Figure 1: Oligonucleotides and poly(L)lysine (pLys) form phase-separated complexes upon mixing. A) 22 nt single-stranded DNA and 50 aa pLys form liquid droplets when mixed at 2.5 mM amine and phosphate concentration. 22 bp double-stranded DNA and 50 aa pLys form solid precipitates when mixed under the same conditions. Imagesmore »
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