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Cutting Through the "Gray Area": An Analysis of the IBC Regulatory Oversight of Applications of CRISPR Technology in Clinical Research.
The advent of clustered regularly interspaced short palindromic repeats (CRISPR) technology has quickly ushered in a new era of gene editing and offered exciting potential for the development of biomedical products. While the breadth of applications for CRISPR encompasses nearly the entire field of medical science, its utilization to produce next-generation CAR T cells stands to benefit most substantially in the short-term. These novel therapeutics are now beginning to enter the clinical trial phase of the numerous approval pipelines, but does the existing regulatory framework have the capability to adequately review and approve the initiation of these new age research endeavors in the clinical setting? This analysis will illuminate the similarity and differences between CRISPR developed interventions and existing techniques used to produce biomedical products from a regulatory perspective in the United States. Further, the "gray area" surrounding CRISPR regulatory oversight will be explored and recommendations will be made to facilitate the elimination of inconsistencies that currently exist in the assessment of this work prior to the initiation of associated clinical trials.
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CRISPR patent ruling leaves license holders scrambling.
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Heteroduplex cleavage assay for screening of probable zygosities resulting from CRISPR mutations in diploid single cell lines.
The most common gene editing methods, such as CRISPR, involve random repair of an induced double-stranded DNA break through the non-homologous end joining (NHEJ) repair pathway, resulting in small insertions/deletions. In diploid cells, these mutations can take on one of three zygosities: monoallelic, diallelic heterozygous, or diallelic homozygous. While many advances have been made in CRISPR delivery systems and gene editing efficiency, little work has been done to streamline detection of gene editing events. The only current method to determine the zygosity of an edited gene in a diploid organism is DNA sequencing, which is costly and time-consuming. Here, we describe the development of a T7 endonuclease I (T7EI)-based heteroduplex cleavage assay, along with statistical models relating the percentage of cleaved DNA to the zygosity of a mutation, that provides a rapid screening step prior to DNA sequencing. By isolating candidates likely to contain the desired zygosity for the edited gene, our screening method can decrease the number of clones requiring DNA sequencing.
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30 years of progress from positional cloning to precision genome editing.
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EU verdict on CRISPR crops dismays scientists.
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Atomic bomb and clustered regularly interspaced short palindromic repeats gene editing technology.
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ZeoR: A Candidate for Assays Testing Enzymatic Activities and Off-Target Effects of Gene-Editing Enzymes.
The target lengths of gene-editing enzymes vary from approximately 20 to 80 nucleotides. The ideal reporter genes should be able to tolerate the insertion of target sequences to differentiate the phenotypic changes between in-frame and frame shift mutations for evaluating the enzymatic activities and off-target effects of gene-editing enzymes such as zinc finger nucleases, transcription activator-like effector nucleases, clustered regularly interspaced short palindromic repeats-associated system. This study demonstrated that the zeocin resistance gene (ZeoR) tolerated the insertion of long targets for a variety of tests using a strategy of inserting tandem repeats of NNT. Four genes such as CCR5, AR, HPV 16E7 early gene, and EGFR applications of ZeoR in quantitatively analyzing the off-targets of CRISPR were successful. The data indicated that evaluating the enzymatic activities and off-target effects with assays employing ZeoR had great potential in facilitating the application of gene editing to human gene therapy.
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CRISPR-S: an active interference element for a rapid and inexpensive selection of genome-edited, transgene-free rice plants.
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First Positive Clinical Data for In Vivo Genome Editing in Humans Opens "New Era of Medicine".
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[Application of clustered regularly interspaced short palindromic repeats- associated protein 9 gene editing technology for treatment of HBV infection].
A lack of effective drugs and technical means to eradicate hepatitis B virus (HBV) is a bottleneck that limits the ability to fully cure HBV infection. Recently, genome-editing technology based on clustered regularly interspaced short palindromic repeats -associated protein 9 is an emerging technique for editing specific gene loci, which can specifically target HBV covalently closed circular DNA, effectively inhibits HBV DNA replication and regulates HBV functional protein expression, and is expected to become a powerful gene therapy tool for the complete eradication of HBV. Considering this, it has become the focus of attention for scholars at home and abroad that how to use clustered regularly interspaced short palindromic repeats -associated protein 9 to accomplish modification of HBV genomes for complete eradication of HBV. This paper summarizes the latest progress based on the latest research results at home and abroad in the application of clustered regularly interspaced short palindromic repeats -associated protein 9 gene editing technology in anti-HBV infection treatment, and expounds its potential and challenges as a radical cure for HBV infection.
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