Chiral sensing combined with nuclease activity assay to track Cas9 dynamics in solution: ROA and CPL study

Monika Halat; Magdalena Klimek-Chodacka; Agnieszka Domagała; Grzegorz Zając; Tomasz Oleszkiewicz; Josef Kapitán; Rafal Baranski

doi:10.1039/D5CC00971E

Chiral sensing combined with nuclease activity assay to track Cas9 dynamics in solution: ROA and CPL study†

Monika Halat,

*^a Magdalena Klimek-Chodacka,

^a Agnieszka Domagała,

^bc Grzegorz Zając,

^b Tomasz Oleszkiewicz,

^a Josef Kapitán

^d and Rafal Baranski

Author affiliations

* Corresponding authors

^a Department of Plant Biology and Biotechnology, Faculty of Biotechnology and Horticulture, University of Agriculture in Krakow, AL. Mickiewicza 21, 31-120 Krakow, Poland
E-mail: m.halat@urk.edu.pl

^b Jagiellonian Centre for Experimental Therapeutics (JCET), Bobrzynskiego 14, 30-348 Krakow, Poland

^c Doctoral School of Exact and Natural Sciences, Jagiellonian University, Prof. S. Łojasiewicza 11, 30-348 Krakow, Poland

^d Department of Optics, Palacký University Olomouc, 17. listopadu 12, Olomouc, Czech Republic

Abstract

Chiroptical studies of the SpyCas9 protein are extremely rare. Nondestructive methods are needed to characterize its active ribonucleoprotein form. Using Raman optical activity (ROA) and circularly polarized luminescence (CPL), we present a new approach to detect key biomolecules involved in CRISPR-Cas technology while preserving their original nucleolytic activity.

This article is part of the themed collection: Chemical Communications HOT articles 2025

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DOI: https://doi.org/10.1039/D5CC00971E
Article type: Communication
Submitted: 21 Feb. 2025
Accepted: 24 Mrt. 2025
First published: 15 Apr. 2025
This article is Open Access

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Chem. Commun., 2025,61, 6905-6908

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Chiral sensing combined with nuclease activity assay to track Cas9 dynamics in solution: ROA and CPL study

M. Halat, M. Klimek-Chodacka, A. Domagała, G. Zając, T. Oleszkiewicz, J. Kapitán and R. Baranski, Chem. Commun., 2025, 61, 6905 DOI: 10.1039/D5CC00971E

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Chemical Communications

Chiral sensing combined with nuclease activity assay to track Cas9 dynamics in solution: ROA and CPL study†

Abstract

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Chiral sensing combined with nuclease activity assay to track Cas9 dynamics in solution: ROA and CPL study

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