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Review
. 2022 May 11;25(6):104389.
doi: 10.1016/j.isci.2022.104389. eCollection 2022 Jun 17.

Integrating CRISPR/Cas systems with programmable DNA nanostructures for delivery and beyond

Petteri Piskunen  1 , Rosalind Latham  2   3 , Christopher E West  2 , Matteo Castronovo  3 , Veikko Linko  1   4
Affiliations
Review

Integrating CRISPR/Cas systems with programmable DNA nanostructures for delivery and beyond

Petteri Piskunen et al. iScience. .

Abstract

Precise genome editing with CRISPR/Cas paves the way for many biochemical, biotechnological, and medical applications, and consequently, it may enable treatment of already known and still-to-be-found genetic diseases. Meanwhile, another rapidly emerging field-structural DNA nanotechnology-provides a customizable and modular platform for accurate positioning of nanoscopic materials, for e.g., biomedical uses. This addressability has just recently been applied in conjunction with the newly developed gene engineering tools to enable impactful, programmable nanotechnological applications. As of yet, self-assembled DNA nanostructures have been mainly employed to enhance and direct the delivery of CRISPR/Cas, but lately the groundwork has also been laid out for other intriguing and complex functions. These recent advances will be described in this perspective.

Keywords: Genetics; Nanostructure; Nanotechnology.

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Conflict of interest statement

The authors declare no competing interests.

Figures

None
Graphical abstract
Figure 1
Figure 1
Diverse CRISPR/Cas functions combined with the DNA nanotechnology toolbox pave the way for new applications CRISPR/Cas tools for genetic editing (genes, bases, primes, and epigenomes), gene regulation and imaging can be integrated with programmable DNA nanostructures to facilitate various predefined functions. Here we discuss especially delivery and targeting but also dynamic operations that are coming increasingly into view. The CRISPR/Cas model is based on the entry CAS9_STAAU from the UniProt database (The UniProt Consortium, 2021).
Figure 2
Figure 2
Delivery systems through CRISPR/Cas-DNA nanostructure fusion (A) β-cyclodextrin cores decorated with linear DNA branches. (B) Tetrahedral DNA nanostructures with vesicle-binding cholesterol linkers and cell-targeting aptamers. (C) Polymer-coated charge-reversible DNA nanoclews. (D) Polymer-coated PCL nanogels cross-linked with DNA linkers. (A) adapted with permission from (Liu et al., 2019); Copyright (2019) American Chemical Society. (B) adapted with permission from (Zhuang et al., 2020); Published (2020) by Oxford Academic Press. (C) adapted with permission from (Sun et al., 2020); Published (2020) by The American Association for the Advancement of Science. (D) adapted with permission from (Huang et al., 2020); Copyright (2020) American Chemical Society.
Figure 3
Figure 3
Advanced applications of CRISPR/Cas-DNA origami nanosystems (A) DNA origami frame for high-speed AFM analysis of Cas cleavage of DNA targets. (B) PAM antennas on DNA origami. (C) Post-processing of DNA origami with CRISPR/Cas. (D) Light-controlled DNA origami-CRISPR/Cas system. (E) Delivery of genes packed into a DNA origami platform. (A) adapted with permission from (Räz et al., 2016); Copyright (2016) American Chemical Society. (B) adapted with permission from (Wang et al., 2020a); Published (2020) by The American Association for the Advancement of Science. (C) adapted with permission from (Xiong et al., 2020); Copyright (2019) John Wiley & Sons. (D) adapted with permission from (Abe et al., 2021); Copyright (2021) by Royal Society of Chemistry. (E) adapted with permission from (Lin-Shiao et al., 2022); Published (2022) Oxford Academic Press.
See this image and copyright information in PMC

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