Generation of the Adenovirus Vector-Mediated CRISPR/Cpf1 System and the Application for Primary Human Hepatocytes Prepared from Humanized Mice with Chimeric Liver

Tsukamoto, Toyohisa; Sakai, Eiko; Iizuka, Shunsuke; Taracena-Gándara, Marcos; Sakurai, Fuminori; Mizuguchi, Hiroyuki

doi:10.1248/bpb.b18-00222

Cited by 23 publications

(17 citation statements)

References 41 publications

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“…Other approaches will edit transgenic silkworms infected with bacterial diseases, fungal diseases and Nosema bombycis according to the characteristics of different silkworm diseases. Currently, the CRISPR family has identified gene editing systems including CRISPR/CPF1 and CRISPR/Cas13, that could edit the DNA, RNA or both DNA and RNA sequences [35,36]. CRISPR/CPF1 and CRISPR/Cas13 systems have smaller nuclease proteins and have more RNA target sequences and lower off-target efficiency.…”

Section: Discussionmentioning

confidence: 99%

A Multiplex CRISPR/Cas9 System for Use as an Anti-BmNPV Therapeutic

Dong¹,

Q²,

Hu³

et al. 2018

Preprint

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Clustered regularly interspaced short palindromic repeats/associated protein 9 nuclease (CRISPR/Cas9) technology guided by a single-guide RNA (sgRNA) has recently opened a new avenue for antiviral therapy. A unique capability of the CRISPR/Cas9 system is multiple genome engineering. However, there are few applications in insect viruses by a single Cas9 enzyme targeting two or more sgRNA at different genomic sites for simultaneous production of multiple DNA breaks. To address the need for multi-gene editing and sustained delivery of multiplex CRISPR/Cas9-based genome engineering tools, we developed a one-vector (pSL1180-Cas9-U6-sgRNA) system to express multiple sgRNA and Cas9 protein to excise Bombyx mori nucleopolyhedrovirus (BmNPV) in insect cells. Here, ie-1, gp64, lef-11, and dnapol genes were screened and identified as multiple sgRNA editing sites according to the BmNPV system infection and DNA replication mechanism. Furthermore, we constructed a multiplex editing vector sgMultiple to efficiently regulate multiplex gene editing steps and inhibit BmNPV replication after viral infection. This is the first report that describes the application of multiplex CRISPR/Cas9 system inhibiting insect virus replication. This multiplex system can significant enable the potential of CRISPR/Cas9-based multiplex genome engineering in transgenic silkworms.

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Section: Discussionmentioning

confidence: 99%

A Multiplex CRISPR/Cas9 System for Use as an Anti-BmNPV Therapeutic

Dong¹,

Q²,

Hu³

et al. 2018

Preprint

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“…Unlike the Cas9 induced blunt end cuts, it creates cuts in a zig zag manner along with requiring a small gRNA [29,30]. Tsukamoto et al have observed reduced mutation rates and enhanced viability during manipulation of human hepatocytes using CRISPR/Cpf1 system [31]. Similarly, other authors have also mentioned advantages in terms of speed, efficiency, specificity and multiplexing option in possible real-time clinical deployment of genome editing [32].…”

Section: Table-1: Various Modifications and Improvements Attempted Over Last Few Years In Crispr/cas Technology A Modified Cas/nucleases mentioning

confidence: 99%

Recent Advancement and Innovations in CRISPR/Cas and CRISPR Related Technologies: A review

Khan¹

2021

BTBP

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CRISPR genome editing technologies have been improving by every passing day. The initial CRISPR/Cas9 technologies, though emerged an improved version of genome editing in competition with TALENS and ZFNs, was nevertheless not free from technical and off-target effects. Technological improvements overtime start addressing issues with original CRISPR/Cas9 technology. The major areas of improvement targeted nucleases and delivery methods. Overtime the nuclease like Cas9 had some modifications like FokI-dCas9, Truncated guide RNAs (tru-gRNAs), Paired Cas9 nickase, Cpf1, Cas6 with Csm/Csr complex and chemically treated Cas9. In terms of delivery methods the improvements came along after almost all methods including viral methods like Recombinant Adeno Associated Viruses (rAAV), Lentivirus (LV), and bacteriophages. The review summarizes various non-viral gene delivery modes including physical methods like electroporation and chemical methods like nano particles, cell-derived membrane vesicles (CMVs) with upgraded developments. The review also compares various modes of delivering CRISPR gene editing machinery.

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“…The most common physical delivery techniques include microinjection and electroporation. Viral delivery vectors include retrovirus, lentivirus, adeno-associated virus (AAV), and adenovirus (Ad) vectors [43]. Viral vectors are especially appropriate vehicles for in vivo works, most commonly used for CRISPR system delivery.…”

Section: Cpf1 Delivery Strategiesmentioning

confidence: 99%

“…Recently, there was a generation of Ad vector-mediated CRISPR/Cpf1 system which proved to be valuable in genome editing in primary human hepatocytes. Despite the lower immunogenicity of AAV vectors than Ad vectors and their more safety for therapeutic applications in humans, AAV vectors have smaller packaging capacity than Ad vectors [43].…”

Section: Cpf1 Delivery Strategiesmentioning

confidence: 99%

“…On the same token, AsCpf1 and LbCpf1 were successfully recruited to correct the disease-related mutations in patient-derived induced pluripotent stem cells (iPS) along with the germline correction in the mouse model [144]. Also, AsCpf1 was recently shown to successfully perform genome replacement with the donor fragment in primary human hepatocytes prepared from humanized mice with a chimeric liver [43]. Note worthily, AsCpf1-gRNA-tRNA system represented potent targeting efficiency within the mammalian cells such as human cell lines, porcine fetal fibroblasts (PFFs) and also the embryos that include rabbit zygotes and porcine parthenogenetic embryos.…”

Section: Application Of Genome Editing Mediated By Crispr Cpf1mentioning

confidence: 99%

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CRISPR Cpf1 proteins: structure, function and implications for genome editing

et al. 2019

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CRISPR and CRISPR-associated (Cas) protein, as components of microbial adaptive immune system, allows biologists to edit genomic DNA in a precise and specific way. CRISPR-Cas systems are classified into two main classes and six types. Cpf1 is a putative type V (class II) CRISPR effector, which can be programmed with a CRISPR RNA to bind and cleave complementary DNA targets. Cpf1 has recently emerged as an alternative for Cas9, due to its distinct features such as the ability to target T-rich motifs, no need for trans-activating crRNA, inducing a staggered double-strand break and potential for both RNA processing and DNA nuclease activity. In this review, we attempt to discuss the evolutionary origins, basic architectures, and molecular mechanisms of Cpf1 family proteins, as well as crRNA designing and delivery strategies. We will also describe the novel Cpf1 variants, which have broadened the versatility and feasibility of this system in genome editing, transcription regulation, epigenetic modulation, and base editing. Finally, we will be reviewing the recent studies on utilization of Cpf1as a molecular tool for genome editing.

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Generation of the Adenovirus Vector-Mediated CRISPR/Cpf1 System and the Application for Primary Human Hepatocytes Prepared from Humanized Mice with Chimeric Liver

Cited by 23 publications

References 41 publications

A Multiplex CRISPR/Cas9 System for Use as an Anti-BmNPV Therapeutic

A Multiplex CRISPR/Cas9 System for Use as an Anti-BmNPV Therapeutic

Recent Advancement and Innovations in CRISPR/Cas and CRISPR Related Technologies: A review

CRISPR Cpf1 proteins: structure, function and implications for genome editing

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