In vivo genome editing in animals using AAV-CRISPR system: applications to translational research of human disease

Lau, Cia-Hin; Suh, Yousin

doi:10.12688/f1000research.11243.1

Cited by 140 publications

(111 citation statements)

References 160 publications

(240 reference statements)

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“…To overcome this problem, a novel strategy was introduced using the smallest Cas9 orthologue derived from Campylobacter jejuni (CjCas9), which was able to successfully reduce VEGFA gene expression in mice with age‐related macular degeneration. In this regard, a new pathway was discovered for the in vivo treatment of age‐related macular degeneration (Table ) …”

Section: Introductionmentioning

confidence: 99%

Creating cell and animal models of human disease by genome editing using CRISPR/Cas9

Zarei

Razban

Hosseini

et al. 2019

The Journal of Gene Medicine

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A set of unique sequences in bacterial genomes, responsible for protecting bacteria against bacteriophages, has recently been used for the genetic manipulation of specific points in the genome. These systems consist of one RNA component and one enzyme component, known as CRISPR (“clustered regularly interspaced short palindromic repeats”) and Cas9, respectively. The present review focuses on the applications of CRISPR/Cas9 technology in the development of cellular and animal models of human disease. Making a desired genetic alteration depends on the design of RNA molecules that guide endonucleases to a favorable genomic location. With the discovery of CRISPR/Cas9 technology, researchers are able to achieve higher levels of accuracy because of its advantages over alternative methods for editing genome, including a simple design, a high targeting efficiency and the ability to create simultaneous alterations in multiple sequences. These factors allow the researchers to apply this technology to creating cellular and animal models of human diseases by knock‐in, knock‐out and Indel mutation strategies, such as for Huntington's disease, cardiovascular disorders and cancers. Optimized CRISPR/Cas9 technology will facilitate access to valuable novel cellular and animal genetic models with respect to the development of innovative drug discovery and gene therapy.

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Section: Introductionmentioning

confidence: 99%

Creating cell and animal models of human disease by genome editing using CRISPR/Cas9

Zarei

Razban

Hosseini

et al. 2019

The Journal of Gene Medicine

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Section: Introductionmentioning

confidence: 99%

“…As such, we have previously shown that rAAV-and zinc-finger nuclease (ZFN)-mediated liver targeting can correct disease phenotypes in neonatal and adult mouse models, a process currently under clinical investigation [18][19][20][21] . Therefore, further development of robust and wide-ranging CRISPR-based technologies for in vivo editing may help to decipher disease mechanisms and offer novel therapeutic options 22,23 .Here we revisited the properties of Streptococcus thermophilus type II-A CRISPR1-Cas9 system, a model nuclease of paramount importance to the entire CRISPR field, and engineered a potent RNA-guided nuclease for both in vitro and in vivo applications. S. thermophilus encodes up to two active type II-A systems (CRISPR1 and CRISPR3) that epitomize the instrumental role played by researchers focusing on the biology of CRISPR-Cas systems in the development of genome-engineering tools 5,24 .…”

mentioning

confidence: 99%

Versatile and robust genome editing with Streptococcus thermophilus CRISPR1-Cas9

Agudelo

Carter

Velimirovic

et al. 2018

Preprint

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The broad spectrum of activities displayed by CRISPR-Cas systems has led to biotechnological innovations that are poised to transform human therapeutics. Therefore, the comprehensive characterization of distinct Cas proteins is highly desirable. Here we expand the repertoire of nucleases for mammalian genome editing using the archetypal Streptococcus thermophilus CRISPR1-Cas9 (St1Cas9). We define functional protospacer adjacent motif (PAM) sequences and variables required for robust and efficient editing in vitro. Expression of holoSt1Cas9 from a single adeno-associated viral (rAAV) vector in the neonatal liver rescued lethality and metabolic defects in a mouse model of hereditary tyrosinemia type I demonstrating effective cleavage activity in vivo. Furthermore, we identified potent anti-CRISPR proteins to regulate the activity of both St1Cas9 and the related type II-A Staphylococcus aureus Cas9 (SaCas9). This work expands the targeting range and versatility of CRISPR-associated enzymes and should encourage studies to determine its structure, genome-wide specificity profile and sgRNA design rules. INTRODUCTIONClustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins form a prokaryotic adaptive immune system and some of its components have been harnessed for robust genome editing 1 . Type II-based editing tools rely on a large multidomain endonuclease, Cas9, guided to its DNA target by an engineered single-guide RNA (sgRNA) chimera 2 (See 3, 4 for a classification of CRISPR-Cas systems). The Cas9-sgRNA binary complex finds its target through recognition of a short sequence called the protospacer adjacent motif (PAM) and subsequent base pairing of the guide RNA with the DNA to generate a specific doublestrand break (DSB) 1,5 . While Streptococcus pyogenes (SpCas9) remains the most widely used Cas9 variant for genome engineering, the diversity of naturally occurring RNA-guided nucleases is astonishing 4 . Hence, Cas9 enzymes from different microbial species can contribute to the expansion of the CRISPR toolset by increasing targeting density, improving activity and specificity as well as easing delivery 1,6 .In principle, engineering complementary CRISPRCas systems from distinct bacterial species should be relatively straightforward, as they have been minimized to only two components. However, many such enzymes were found inactive in human cells despite being accurately reprogrammed for DNA binding and cleavage in vitro [7][8][9][10] . Nevertheless, the full potential of selected enzymes can be unleashed using machine learning to establish sgRNA design rules 11,12 .Perhaps the most striking example of the value of alternative Cas9 enzymes is the implementation of the type II-A Cas9 from Staphylococcus aureus (SaCas9) for in vivo editing using recombinant adeno-associated virus (rAAV) vectors 7,13, 14 . More recently, Campylobacter jejuni and Neisseria meningitidis Cas9s from the type II-C 15 CRISPRCas systems have been added to this repertoire 16,17 .In vivo genome edi...

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“…5 Nonvirally-mediated transient expression of CRISPR components in the retina may reduce safety concerns, although viral delivery systems based on AAV represent the most efficient and safe tools for gene delivery to the retina. Indeed genome editing using the AAV-CRISPR system has been widely reported as efficient, safe, and precise in more than 30 published studies in mouse models 6 of diseases associated with the eyes, muscle, liver, heart, and lung. Despite the great potential of AAV vectors, their relatively small packaging capacity represents a limitation for delivering the widely used Streptococcus pyogenes Cas9 (SpCas9) together with guide RNAs (gRNAs) and large transgenes.…”

mentioning

confidence: 99%

AAV-CRISPR Persistence in the Eye of the Beholder

Recchia

2019

Molecular Therapy

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In vivo genome editing in animals using AAV-CRISPR system: applications to translational research of human disease

Cited by 140 publications

References 160 publications

Creating cell and animal models of human disease by genome editing using CRISPR/Cas9

Creating cell and animal models of human disease by genome editing using CRISPR/Cas9

Versatile and robust genome editing with Streptococcus thermophilus CRISPR1-Cas9

AAV-CRISPR Persistence in the Eye of the Beholder

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