Cas9-mediated targeting of viral RNA in eukaryotic cells

Price, Aryn A; Sampson, Timothy R.; Ratner, Hannah K.; Grakoui, Arash; Weiss, David S.

doi:10.1073/pnas.1422340112

Cited by 234 publications

(160 citation statements)

References 36 publications

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“…It will be interesting to see whether sgRNA-Cas9 nickases can be programmed to cut singlestranded RNAs by RNA interference (RNAi). It has been shown that Cas9 endonuclease from Francisella novicida (FnCas9) in complex with sgRNA can be used to target (+) strand ss-viral RNA and hepatitis C virus RNA and inhibit viral replication (93). Thermus thermopilus type III-B CRISPR-Cas systems exclusively target ssRNA for degradation and not DNA sequences that are complementary to the crRNA [reviewed in (76)].…”

Section: With Nicking Enzymes Guided By Rnamentioning

confidence: 99%

Sequence-specific DNA nicking endonucleases

2015

Biomolecular Concepts

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A group of small HNH nicking endonucleases (NEases) was discovered recently from phage or prophage genomes that nick double-stranded DNA sites ranging from 3 to 5 bp in the presence of Mg2+ or Mn2+. The cosN site of phage HK97 contains a gp74 nicking site AC↑CGC, which is similar to AC↑CGR (R=A/G) of N.ϕGamma encoded by Bacillus phage Gamma. A minimal nicking domain of 76 amino acid residues from N.ϕGamma could be fused to other DNA binding partners to generate chimeric NEases with new specificities. The biological roles of a few small HNH endonucleases (HNHE, gp74 of HK97, gp37 of ϕSLT, ϕ12 HNHE) have been demonstrated in phage and pathogenicity island DNA packaging. Another group of NEases with 3- to 7-bp specificities are either natural components of restriction systems or engineered from type IIS restriction endonucleases. A phage group I intron-encoded HNH homing endonucleases, I-PfoP3I was found to nick DNA sites of 14-16 bp. I-TslI encoded by T7-like ΦI appeared to nick DNA sites with a 9-bp core sequence. DNA nicking and labeling have been applied to optical mapping to aid genome sequence assembly and detection of large insertion/deletion mutations in genomic DNA of cancer cells. Nicking enzyme-mediated amplification reaction has been applied to rapid diagnostic testing of influenza A and B in clinical setting and for construction of DNA-based Boolean logic gates. The clustered regularly interspaced short palindromic repeats-ribonucleoprotein complex consisting of engineered Cas9 nickases in conjunction with tracerRNA:crRNA or a single-guide RNA have been successfully used in genome modifications.

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Section: With Nicking Enzymes Guided By Rnamentioning

confidence: 99%

Sequence-specific DNA nicking endonucleases

2015

Biomolecular Concepts

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“…FnCas9 was recently used to target the genome of the positive sense single-stranded RNA (+ssRNA) virus, hepatitis C virus (HCV), as a proof of principle [102]. HCV is an important human pathogen that can cause liver fibrosis, cirrhosis, and may lead to hepatocellular carcinoma [103].…”

Section: Cas9 Targeting Of Rnamentioning

confidence: 99%

Harnessing the Prokaryotic Adaptive Immune System as a Eukaryotic Antiviral Defense

Price

Grakoui

Weiss

2016

Trends in Microbiology

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Clustered, regularly interspaced, short palindromic repeats - CRISPR associated (CRISPR-Cas) systems are sequence specific RNA-directed endonuclease complexes that bind and cleave nucleic acids. These systems evolved within prokaryotes as adaptive immune defenses to target and degrade nucleic acids derived from bacteriophages and other foreign genetic elements. The antiviral function of these systems has now been exploited to combat eukaryotic viruses throughout the viral life cycle. Here we discuss current advances in CRISPR-Cas9 technology as a eukaryotic antiviral defense.

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“…Similarly, in human induced pluripotent stem cells (iPSCs) derived from a myeloproliferative neoplasm, Cas9 was used to repair the oncogenic mutation (Smith et al 2015), and mutations in the crygc gene that is responsible for cataracts were repaired in mouse zygotes and spermatogonial stem cells (Ren et al 2013; Wu et al 2015). Additionally, HIV proviruses have been removed from infected cells using Cas9-directed cleavage, and hepatitis B and hepatitis C viruses have been targeted, perhaps providing a framework for future antiviral therapeutics (Hu et al 2014; Lin et al 2014a; Kennedy et al 2015; Liao et al 2015; Price et al 2015). Such repair has not been limited to tissue culture studies ex vivo.…”

Section: Use Of Cas9 For Genome Engineeringmentioning

confidence: 99%

Overview of CRISPR–Cas9 Biology

Ratner

Sampson

Weiss

2016

Cold Spring Harb Protoc

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Prokaryotes use diverse strategies to improve fitness in the face of different environmental threats and stresses, including those posed by mobile genetic elements (e.g., bacteriophages and plasmids). To defend against these elements, many bacteria and archaea use elegant, RNA-directed, nucleic acid– targeting adaptive restriction machineries called CRISPR–Cas (CRISPR-associated) systems. While providing an effective defense against foreign genetic elements, these systems have also been observed to play critical roles in regulating bacterial physiology during environmental stress. Increasingly, CRISPR–Cas systems, in particular the Type II systems containing the Cas9 endonuclease, have been exploited for their ability to bind desired nucleic acid sequences, as well as direct sequence-specific cleavage of their targets. Cas9-mediated genome engineering is transcending biological research as a versatile and portable platform for manipulating genetic content in myriad systems. Here, we present a systematic overview of CRISPR–Cas history and biology, highlighting the revolutionary tools derived from these systems, which greatly expand the molecular biologists' toolkit.

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Cas9-mediated targeting of viral RNA in eukaryotic cells

Cited by 234 publications

References 36 publications

Sequence-specific DNA nicking endonucleases

Sequence-specific DNA nicking endonucleases

Harnessing the Prokaryotic Adaptive Immune System as a Eukaryotic Antiviral Defense

Overview of CRISPR–Cas9 Biology

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