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Comments on the CRISPR Gene-edited babies' case.
Twin girls were born in China in November 2018 to an HIV-positive father and an HIV-negative mother, through in vitro fertilisation in combination with CRISPR-Cas9 genome editing which altered their CCR5 genes. An investigation found that regulations were broken, and documents were forged. The government needs to address various regulatory defects, improve oversight of research, and implement institutional policies on conflict of interest.
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Multiplex Generation, Tracking, and Functional Screening of Substitution Mutants Using a CRISPR/Retron System.
We developed a clustered regularly interspaced short palindromic repeats (CRISPR)/retron system for multiplexed generation of substitution mutations by coutilization of a retron system that continuously expresses donor DNA and a CRISPR/Cas9 cassette that induces cleavage at target genomic loci. Our system efficiently introduces substitution mutation in the Escherichia coli genome in a high-throughput manner. These substitution mutations can be tracked by analysis of retron plasmid sequences without laborious amplification of individual edited loci. We demonstrated that our CRISPR/retron system can introduce thousands of mutations in a single experiment and be used for screening phenotypes related to chemical responses or fitness changes. We expect that our system could facilitate genome-scale substitution screenings.
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CRISPR/Cas technology promotes the various application of Dunaliella salina system.
Dunaliella salina (D. salina) has been widely applied in various fields because of its inherent advantages, such as the study of halotolerant mechanism, wastewater treatment, recombinant proteins expression, biofuel production, preparation of natural materials, and others. However, owing to the existence of low yield or in the laboratory exploration stage, D. salina system has been greatly restricted for practical production of various components. In past decade, significant progresses have been achieved for research of D. salina in these fields. Among them, D. salina as a novel expression system demonstrated a bright prospect, especially for large-scale production of foreign proteins, like the vaccines, antibodies, and other therapeutic proteins. Due to the low efficiency, application of traditional regulation tools is also greatly limited for exploration of D. salina system. The emergence of the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system offers a precise editing tool to overcome the obstacles of D. salina system. This review not only comprehensively summarizes the recent progresses of D. salina in domain of gene engineering but also gives a deep analysis of problems and deficiencies in different fields of D. salina. Moreover, further prospects of CRISPR/Cas system and its significant challenges have been discussed in various aspects of D. salina. It provides a great referencing value for speeding up the maturity of D. salina system, and also supplies practical guiding significance to expand the new application fields for D. salina. KEY POINTS: • The review provides recent research progresses of various applications of D. salina. • The problems and deficiencies in different fields of D. salina were deeply analyzed. • The further prospects of CRISPR/Cas technology in D. salina system were predicted. • CRISPR/Cas system will promote the new application fields and maturity for D. salina.
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Commentary - CRISPR-based techniques: Cas9, Cas13 and their applications in the era of COVID-19.
The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9 (CRISPR-associated protein 9) system enables scientists to edit diverse genome types with relative ease, with the aim - in the near future - to prevent future human beings from developing genetic diseases. The new opportunities arising from the system are broad-ranging and revolutionary, but such prospects have also been the cause for alarm throughout the international scientific community. The authors have laid out a review of the trials carried out so far in terms of genome editing, for the ultimate purpose of weighing implications and criticisms. We feel that possible valuable alternatives, such as induced pluripotent stem cells should not be overlooked.
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GOANA: A Universal High-Throughput Web Service for Assessing and Comparing the Outcome and Efficiency of Genome Editing Experiments.
The increased development of functionally diverse and highly specialized genome editors has created the need for comparative analytics tools that are able to profile the mutational outcomes, particularly rare and complex outcomes, to assess the editor's applicability to different domains. To address this need, we have developed Generalizable On-target activity ANAlyzer (GOANA), a high-throughput web-based software for determining editing efficiency and cataloguing rare outcomes from next-generation sequencing data. GOANA calculates mutation frequency and outcomes relative to a supplied control sample. It is scalable to thousands of target sites across the entire genome and is 4,000% faster than CRISPResso2. Mutations are reported on a "per-read" level rather than individually, enabling the identification of co-occurring mutations. GOANA is editor agnostic and can be applied to data generated from any targeted editing experiment, including base editors. Requiring only that control and treated reads are aligned to the same reference, GOANA can handle data from any library preparation method, including pooled amplicon and whole-genome sequencing. As a proof of principle, we analyze two large data sets of CRISPR-Cas9 and CRISPR-Cas12a editing, demonstrating the power of GOANA and highlighting several key differences between the two enzymes. GOANA is available for use at https://gt-scan.csiro.au/goana/ and as a command line tool from https://github.com/BauerLab/GOANA.
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CRISPR Hits Home in a First-in-Human Study.
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Too Much Compromise in Today's CRISPR Pipelines.
CRISPR-based editing is a revolutionary tool for genome engineering and discovery. The pace of innovation in this young field is truly astonishing, and yet significant gaps remain in the set of capabilities offered. In particular, scalable (massively parallel and multiplex) editing is only available for a small variety of edit types, placing fundamental limits on the kinds of studies researchers can perform. Additional advancements are needed to realize the full potential of the technology.
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Engineering human knock-in organoids.
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CRISPR-based gene drive in agriculture will face technical and governance challenges.
Comment on “Agricultural pest control with CRISPR‐based gene drive: time for public debate” by Courtier‐Orgogozo et al. [Image: see text]
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Letter to the Editor: The World's First CRISPR Tomato Launched to a Japanese Market: The Social-Economic Impact of its Implementation on Crop Genome Editing.
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