Spatial control of in vivo CRISPR–Cas9 genome editing via nanomagnets

Abstract

The potential of CRISPR–Cas9-based therapeutic genome editing is hampered by difficulties in the control of the in vivo activity of CRISPR–Cas9. To minimize any genotoxicity, precise activation of CRISPR–Cas9 in the target tissue is desirable. Here, we show that, by complexing magnetic nanoparticles (MNPs) with recombinant baculoviral vectors (BVs), CRISPR–Cas9-mediated genome editing can be activated locally in vivo via a magnetic field. BV was chosen for in vivo gene delivery because of its large loading capacity and its ability to locally overcome systemic inactivation by the complement system. We demonstrate that a locally applied magnetic field can enhance the cellular entry of MNP-BVs, thereby avoiding BV inactivation and causing a transient transgene expression in the target tissue. Because BVs are inactivated elsewhere, gene delivery and in vivo genome editing via MNP-BVs are tissue-specific.

Beyond the qualitative description of complex magnetic nanoparticle arrays using FORC measurement

“…Advancement of nanotechnology has extensively expedited the emergence of novel magnetic nanostructures, such as magnetic nanowires (MNWs), in various research areas, including medical treatment [1][2][3][4][5], environmental science [6,7], and quantum devices [8][9][10][11][12]. These magnetic nanostructures have opened numerous opportunities for scientists in different disciplines such as nanomedicine, molecular biology [13][14][15][16], applied physics, and nanostructured materials [17][18][19][20][21][22]. In all of these applications, it is crucial to know the characteristics of the magnetic nanostructures, which may inhibit or enhance their use depending on the application.…”

Engineering a CRISPR Interference System To Repress a Class 1 Integron in Escherichia coli

“…For instance, based on experimentally validated sgRNA R3, a vehicle encompassing sgRNA R3 and the dCas9 could be prepared and delivered to desired settings and subsequently respond to cues such as pH (30), light (29,32), or magnetism (33) for eliciting a CRISPRi system. In fact, CRISPR delivery systems have been under construction recently (29,30,32,33). For instance, one group from Nanjing University attempted to cure a tumor by a near-infrared (NIR) light-responsive nano carrier of CRISPR-Cas9 (32).…”

“…For instance, one group from Nanjing University attempted to cure a tumor by a near-infrared (NIR) light-responsive nano carrier of CRISPR-Cas9 (32). Shortly afterwards, another group from Rice University reported spatial control of CRISPR editing through an artificial magnetic-field-driven baculoviral vector (33). These interdisciplinary studies are broadening the applications of the CRISPR system (34).…”

“…These interdisciplinary studies are broadening the applications of the CRISPR system (34). In addition to the aforementioned nanoparticles (32,33), the CRISPR system can be delivered by conjugative plasmids (35,36), phagemids (35), mobile genomic islands (37), or transposons (38,39). Given this information, we envision that the CRISPR system may be fused to an integron, resulting in a chimeric integron which is mobile.…”

Heat-triggered remote control of CRISPR-dCas9 for tunable transcriptional modulation

“…Toward this end, several inducible systems have been developed to provide the ability to modulate the activity of Cas9 and its variants in living animals [4][5] . These include Cas9 systems that rely on chemical triggers using small molecule drugs, such as rapamycin and tamoxifen [6][7][8] , to activate and tune Cas9 activity by defined doses and at specific points in time.…”