CRISPR/Cas9 produces anti-hepatitis B virus effect in hepatoma cells and transgenic mouse

Zhu, Wei; Xie, Kun; Xu, Y.; Wang, Le; Chen, Kaiming; Zhang, Longzhen; Fang, Jianmin

doi:10.1016/j.virusres.2016.04.003

Cited by 54 publications

(44 citation statements)

References 36 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%

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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%

“…However, several studies have demonstrated that HBV cccDNA could be cleaved by CRISPR/Cas9 system. They have shown that targeting different ORFs using a single sgRNA might be effective elimination cccDNA but give better results if multiple sgRNAs target different loci …”

Section: Introductionmentioning

confidence: 99%

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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“…Viral vectors used for ex vivo and in vivo delivery of nuclease-encoding genes include adeno-associated virus 29,30,31,32,33 , adenovirus 34,35,36,37 , and lentivirus 38,39,40,41,42 . Additionally, non-viral delivery systems, lipid-based nanoparticles 24,25,43,44,45,46,47,48 , polymeric nanoparticles 49,50,51,52 , and cell-penetrating peptides 26,27 have been investigated.…”

Section: Introductionmentioning

confidence: 99%

Therapeutic gene editing: delivery and regulatory perspectives

et al. 2017

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Gene-editing technology is an emerging therapeutic modality for manipulating the eukaryotic genome by using target-sequence-specific engineered nucleases. Because of the exceptional advantages that gene-editing technology offers in facilitating the accurate correction of sequences in a genome, gene editing-based therapy is being aggressively developed as a next-generation therapeutic approach to treat a wide range of diseases. However, strategies for precise engineering and delivery of gene-editing nucleases, including zinc finger nucleases, transcription activator-like effector nuclease, and CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats-associated nuclease Cas9), present major obstacles to the development of gene-editing therapies, as with other gene-targeting therapeutics. Currently, viral and non-viral vectors are being studied for the delivery of these nucleases into cells in the form of DNA, mRNA, or proteins. Clinical trials are already ongoing, and in vivo studies are actively investigating the applicability of CRISPR/Cas9 techniques. However, the concept of correcting the genome poses major concerns from a regulatory perspective, especially in terms of safety. This review addresses current research trends and delivery strategies for gene editing-based therapeutics in non-clinical and clinical settings and considers the associated regulatory issues.

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“…Derived from the bacterial type II CRISPR/Cas system, the Cas9 nuclease can be directed by a programmable single guide RNA (sgRNA) to induce double strand breaks at specific loci, triggering the NHEJ or HDR repair pathways to yield desired sequence modifications (18 -21). Using CRISPR, a variety of simple and sophisticated genomic modification schemes have been generated in mouse models, including precise sequence replacement (22)(23)(24), genome insertions/deletions (25,26), and chromosomal translocations (27). The standard practice for CRISPR editing in mice relies on microinjection of Cas9 DNA/mRNA and sgRNAs into one-cell zygotes (15,28), a process that remains laborious, costly, and low throughput with considerable technical barriers.…”

mentioning

confidence: 99%

Highly Efficient Mouse Genome Editing by CRISPR Ribonucleoprotein Electroporation of Zygotes

Chen

Lee

et al. 2016

Journal of Biological Chemistry

271

243

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The CRISPR/Cas9 system has been employed to efficiently edit the genomes of diverse model organisms. CRISPR-mediated mouse genome editing is typically accomplished by microinjection of Cas9 DNA/RNA and single guide RNA (sgRNA) into zygotes to generate modified animals in one step. However, microinjection is a technically demanding, labor-intensive, and costly procedure with poor embryo viability. Here, we describe a simple and economic electroporation-based strategy to deliver Cas9/sgRNA ribonucleoproteins into mouse zygotes with 100% efficiency for in vivo genome editing. Our methodology, designated as CRISPR RNP Electroporation of Zygotes (CRISPR-EZ), enables highly efficient and high-throughput genome editing in vivo, with a significant improvement in embryo viability compared with microinjection. Using CRISPR-EZ, we generated a variety of editing schemes in mouse embryos, including indel (insertion/deletion) mutations, point mutations, large deletions, and small insertions. In a proof-of-principle experiment, we used CRISPR-EZ to target the tyrosinase (Tyr) gene, achieving 88% bi-allelic editing and 42% homology-directed repairmediated precise sequence modification in live mice. Taken together, CRISPR-EZ is simple, economic, high throughput, and highly efficient with the potential to replace microinjection for in vivo genome editing in mice and possibly in other mammals.

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CRISPR/Cas9 produces anti-hepatitis B virus effect in hepatoma cells and transgenic mouse

Cited by 54 publications

References 36 publications

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

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

Therapeutic gene editing: delivery and regulatory perspectives

Highly Efficient Mouse Genome Editing by CRISPR Ribonucleoprotein Electroporation of Zygotes

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