Hierarchical Self-uncloaking CRISPR-Cas13a-customized RNA Nanococoons for Spatial-controlled Genome Editing and Precise Cancer Therapy

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2022 May 18

PMID 35584220

Authors

Ningke Fan

Xintong Bian

Meng Li

Junman Chen

Haiping Wu

Qiling Peng

Huijie Bai

Wenqian Cheng

Liangsheng Kong

Shijia Ding

Siqiao Li

Wei Cheng

Affiliations

Soon will be listed here.

Abstract

CRISPR-Cas13a holds enormous potential for developing precise RNA editing. However, spatial manipulation of CRISPR-Cas13a activity remains a daunting challenge for elaborately regulating localized RNase function. Here, we designed hierarchical self-uncloaking CRISPR-Cas13a-customized RNA nanococoons (RNCOs-D), featuring tumor-specific recognition and spatial-controlled activation of Cas13a, for precise cancer synergistic therapy. RNCOs-D consists of programmable RNA nanosponges (RNSs) capable of targeted delivery and caging chemotherapeutic drug, and nanocapsules (NCs) anchored on RNSs for cloaking Cas13a/crRNA ribonucleoprotein (Cas13a RNP) activity. The acidic endo/lysosomal microenvironment stimulates the outer decomposition of NCs with concomitant Cas13a RNP activity revitalization, while the inner disassembly through trans-cleavage of RNSs initiated by cis-recognition and cleavage of EGFR variant III (EGFRvIII) mRNA. RNCOs-D demonstrates the effective EGFRvIII mRNA silencing for synergistic therapy of glioblastoma cancer cells in vitro and in vivo. The engineering of RNSs, together with efficient Cas13a activity regulation, holds immense prospect for multimodal and synergistic cancer therapy.

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References

Kushawah G, Hernandez-Huertas L, Abugattas-Nunez Del Prado J, Martinez-Morales J, DeVore M, Hassan H . CRISPR-Cas13d Induces Efficient mRNA Knockdown in Animal Embryos. Dev Cell. 2020; 54(6):805-817.e7. DOI: 10.1016/j.devcel.2020.07.013. View

Pecot C, Calin G, Coleman R, Lopez-Berestein G, Sood A . RNA interference in the clinic: challenges and future directions. Nat Rev Cancer. 2010; 11(1):59-67. PMC: 3199132. DOI: 10.1038/nrc2966. View

Yue H, Huang R, Shan Y, Xing D . Delivery of Cas13a/crRNA by self-degradable black phosphorus nanosheets to specifically inhibit Mcl-1 for breast cancer therapy. J Mater Chem B. 2020; 8(48):11096-11106. DOI: 10.1039/d0tb01914c. View

Afonin K, Grabow W, Walker F, Bindewald E, Dobrovolskaia M, Shapiro B . Design and self-assembly of siRNA-functionalized RNA nanoparticles for use in automated nanomedicine. Nat Protoc. 2011; 6(12):2022-34. PMC: 3498981. DOI: 10.1038/nprot.2011.418. View

Jasinski D, Haque F, Binzel D, Guo P . Advancement of the Emerging Field of RNA Nanotechnology. ACS Nano. 2017; 11(2):1142-1164. PMC: 5333189. DOI: 10.1021/acsnano.6b05737. View

Lv Y, Hu R, Zhu G, Zhang X, Mei L, Liu Q . Preparation and biomedical applications of programmable and multifunctional DNA nanoflowers. Nat Protoc. 2015; 10(10):1508-24. PMC: 4927333. DOI: 10.1038/nprot.2015.078. View

East-Seletsky A, OConnell M, Knight S, Burstein D, Cate J, Tjian R . Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016; 538(7624):270-273. PMC: 5576363. DOI: 10.1038/nature19802. View

Ni Q, Zhang F, Zhang Y, Zhu G, Wang Z, Teng Z . In Situ shRNA Synthesis on DNA-Polylactide Nanoparticles to Treat Multidrug Resistant Breast Cancer. Adv Mater. 2018; 30(10). DOI: 10.1002/adma.201705737. View

Narita Y, Nagane M, Mishima K, Huang H, Furnari F, Cavenee W . Mutant epidermal growth factor receptor signaling down-regulates p27 through activation of the phosphatidylinositol 3-kinase/Akt pathway in glioblastomas. Cancer Res. 2002; 62(22):6764-9. View

10.

Meeske A, Jia N, Cassel A, Kozlova A, Liao J, Wiedmann M . A phage-encoded anti-CRISPR enables complete evasion of type VI-A CRISPR-Cas immunity. Science. 2020; 369(6499):54-59. PMC: 7975689. DOI: 10.1126/science.abb6151. View

11.

Qin X, Li Y . Strategies To Design and Synthesize Polymer-Based Stimuli-Responsive Drug-Delivery Nanosystems. Chembiochem. 2020; 21(9):1236-1253. DOI: 10.1002/cbic.201900550. View

12.

Wu T, Liu J, Liu M, Liu S, Zhao S, Tian R . A Nanobody-Conjugated DNA Nanoplatform for Targeted Platinum-Drug Delivery. Angew Chem Int Ed Engl. 2019; 58(40):14224-14228. DOI: 10.1002/anie.201909345. View

13.

Park Y, Kim H, Lee J . Self-assembled DNA-Guided RNA Nanovector via Step-wise Dual Enzyme Polymerization (SDEP) for Carrier-free siRNA Delivery. ACS Biomater Sci Eng. 2021; 2(4):616-624. DOI: 10.1021/acsbiomaterials.5b00554. View

14.

Verbeke S, Perret R, Chaire V, Richard E, Velasco V, Giles F . GSK3-beta as a candidate therapeutic target in soft tissue sarcomas. J Hematol Oncol. 2021; 14(1):202. PMC: 8641200. DOI: 10.1186/s13045-021-01215-x. View

15.

Lee J, Hong J, Bonner D, Poon Z, Hammond P . Self-assembled RNA interference microsponges for efficient siRNA delivery. Nat Mater. 2012; 11(4):316-22. PMC: 3965374. DOI: 10.1038/nmat3253. View

16.

Zhang Z, Wang Q, Liu Q, Zheng Y, Zheng C, Yi K . Dual-Locking Nanoparticles Disrupt the PD-1/PD-L1 Pathway for Efficient Cancer Immunotherapy. Adv Mater. 2019; 31(51):e1905751. DOI: 10.1002/adma.201905751. View

17.

Hanahan D, Weinberg R . Hallmarks of cancer: the next generation. Cell. 2011; 144(5):646-74. DOI: 10.1016/j.cell.2011.02.013. View

18.

Mohsen M, Kool E . The Discovery of Rolling Circle Amplification and Rolling Circle Transcription. Acc Chem Res. 2016; 49(11):2540-2550. PMC: 5568012. DOI: 10.1021/acs.accounts.6b00417. View

19.

Chen X, Rechavi O . Plant and animal small RNA communications between cells and organisms. Nat Rev Mol Cell Biol. 2021; 23(3):185-203. PMC: 9208737. DOI: 10.1038/s41580-021-00425-y. View

20.

Abudayyeh O, Gootenberg J, Konermann S, Joung J, Slaymaker I, Cox D . C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 2016; 353(6299):aaf5573. PMC: 5127784. DOI: 10.1126/science.aaf5573. View