A non-viral CRISPR/Cas9 delivery system for therapeutically targeting HBV DNA and pcsk9 in vivo

Jiang, Chao; Mei, Miao; Li, Bin; Zhu, Xiurui; Zu, Wenhong; Tian, Yujie; Wang, Qiannan; Guo, Yong; Dong, Yizhou; Tan, Xu

doi:10.1038/cr.2017.16

Cited by 271 publications

(217 citation statements)

References 15 publications

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“…Intriguingly, they also have shown that the combination of the sgRNAs was more efficient in suppressing HBV protein in comparison to individual sgRNAs . In agreement with previous study, several other groups have shown significant drop in the total level of HBV DNA and HBV proteins via anti HBV sgRNAs against different regions of HBV genome . As indicated before, elimination of HBV cccDNA in the hepatocyte nuclei seems to be impossible with current therapies; therefore, the removal of cccDNA is an important aim in eradicating of HBV infection.…”

Section: Introductionsupporting

confidence: 84%

Harnessing CRISPR/Cas 9 System for manipulation of DNA virus genome

Ebrahimi

Teimoori

Khanbabaei

et al. 2018

Reviews in Medical Virology

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Summary The recent development of the Clustered Regularly Interspaced Palindromic Repeat (CRISPR)/CRISPR‐associated protein 9 (Cas9) system, a genome editing system, has many potential applications in virology. The possibility of introducing site specific breaks has provided new possibilities to precisely manipulate viral genomics. Here, we provide diagrams to summarize the steps involved in the process. We also systematically review recent applications of the CRISPR/Cas9 system for manipulation of DNA virus genomics and discuss the therapeutic potential of the system to treat viral diseases.

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Section: Introductionsupporting

confidence: 84%

Harnessing CRISPR/Cas 9 System for manipulation of DNA virus genome

Ebrahimi

Teimoori

Khanbabaei

et al. 2018

Reviews in Medical Virology

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“…Nonviral delivery methods such as membrane deformation7 or hydrodynamic injection8, 9 have been used to deliver plasmid DNA expressing Cas9 and sgRNA, but the possible damage of the target cells and/or the unsatisfactory delivery efficiency compromise the gene editing efficacy. The delivery of in vitro‐transcribed mRNA of Cas9 and sgRNA,10, 11 although promising in offering an alternative to DNA delivery, also faces challenges such as immunogenicity and the poor stability of the RNA 12. The delivery of the complex of Cas9 protein and sgRNA, although can realize targeting gene editing, still faces the problem of the instability of the RNA 13, 14, 15.…”

Section: Introductionmentioning

confidence: 99%

Genome Editing for Cancer Therapy: Delivery of Cas9 Protein/sgRNA Plasmid via a Gold Nanocluster/Lipid Core–Shell Nanocarrier

et al. 2017

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The type II bacterial clustered, regularly interspaced, short palindromic repeats (CRISPR)‐Cas9 (CRISPR‐associated protein) system (CRISPR‐Cas9) is a powerful toolbox for gene‐editing, however, the nonviral delivery of CRISPR‐Cas9 to cells or tissues remains a key challenge. This paper reports a strategy to deliver Cas9 protein and single guide RNA (sgRNA) plasmid by a nanocarrier with a core of gold nanoclusters (GNs) and a shell of lipids. By modifying the GNs with HIV‐1‐transactivator of transcription peptide, the cargo (Cas9/sgRNA) can be delivered into cell nuclei. This strategy is utilized to treat melanoma by designing sgRNA targeting Polo‐like kinase‐1 (Plk1) of the tumor. The nanoparticle (polyethylene glycol‐lipid/GNs/Cas9 protein/sgPlk1 plasmid, LGCP) leads to >70% down‐regulation of Plk1 protein expression of A375 cells in vitro. Moreover, the LGCP suppresses melanoma progress by 75% on mice. Thus, this strategy can deliver protein‐nucleic acid hybrid agents for gene therapy.

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“…By contrast, all previous systemically administered nanoparticle gene editing has occurred preferentially in hepatocytes. 46,48–50 This is the first report of sgRNA-mediated in vivo editing in hepatic endothelial cells.…”

Section: Resultsmentioning

confidence: 87%

Analyzing 2000 in Vivo Drug Delivery Data Points Reveals Cholesterol Structure Impacts Nanoparticle Delivery

et al. 2018

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Lipid nanoparticles (LNPs) are formulated using unmodified cholesterol. However, cholesterol is naturally esterified and oxidized in vivo, and these cholesterol variants are differentially trafficked in vivo via lipoproteins including LDL and VLDL. We hypothesized that incorporating the same cholesterol variants into LNPs - which can be structurally similar to LDL and VLDL – would alter nanoparticle targeting in vivo. To test this hypothesis, we quantified how >100 LNPs made with 6 cholesterol variants delivered DNA barcodes to 18 cell types in wildtype, LDL R−/−, and VLDLR−/− mice that were both age-matched and female. By analyzing ~2,000 in vivo drug delivery data points, we found that LNPs formulated with esterified cholesterol delivered nucleic acids more efficiently than LNPs formulated with regular or oxidized cholesterol when compared across all tested cell types in the mouse. We also identified an LNP containing cholesteryl oleate that efficiently delivered siRNA and sgRNA to liver endothelial cells in vivo. Delivery was as - or more - efficient than the same LNP made with unmodified cholesterol. Moreover, delivery to liver endothelial cells was 3X more efficient than delivery to hepatocytes, distinguishing this oleate LNP from hepatocyte-targeting LNPs. RNA delivery can be improved by rationally selecting cholesterol variants, allowing optimization of nanoparticle targeting.

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A non-viral CRISPR/Cas9 delivery system for therapeutically targeting HBV DNA and pcsk9 in vivo

Cited by 271 publications

References 15 publications

Harnessing CRISPR/Cas 9 System for manipulation of DNA virus genome

Harnessing CRISPR/Cas 9 System for manipulation of DNA virus genome

Genome Editing for Cancer Therapy: Delivery of Cas9 Protein/sgRNA Plasmid via a Gold Nanocluster/Lipid Core–Shell Nanocarrier

Analyzing 2000 in Vivo Drug Delivery Data Points Reveals Cholesterol Structure Impacts Nanoparticle Delivery

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