CRISPR Journal最新文献

A "Bare Hope of A Result": The Second CRISPR Patent Appeal. “结果的一线希望”：第二次CRISPR专利上诉。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2025-07-17 DOI: 10.1177/25731599251361362

Jacob S Sherkow

引用次数: 0

Bacterial Expression System with Deep Repression and Activation via CRISPR-Cas9. 通过CRISPR-Cas9深层抑制和激活的细菌表达系统。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2025-07-14 DOI: 10.1177/25731599251358852

Valentin A Manuvera, Pavel A Bobrovsky, Daria D Kharlampieva, Ekaterina N Grafskaia, Ksenia A Brovina, Maria Y Serebrennikova, Vassili N Lazarev

{"title":"Bacterial Expression System with Deep Repression and Activation via CRISPR-Cas9.","authors":"Valentin A Manuvera, Pavel A Bobrovsky, Daria D Kharlampieva, Ekaterina N Grafskaia, Ksenia A Brovina, Maria Y Serebrennikova, Vassili N Lazarev","doi":"10.1177/25731599251358852","DOIUrl":"https://doi.org/10.1177/25731599251358852","url":null,"abstract":"Incomplete repression of recombinant genes encoding toxic polypeptides can suppress cell growth even in the absence of a transcription inducer. To address this issue, we developed a CRISPR-Cas9-based genome editing approach that directly modifies the plasmid encoding the toxic peptide during Escherichia coli cultivation. The constructed plasmids contained a transcription terminator between the promoter and coding region, preventing full gene expression through abortive transcription. Upon CRISPR-Cas9 activation, this region is excised, thus restoring the functional gene. To implement this approach, we modified widely used pET-series expression plasmids by adding extra terminators in the 5'-untranslated region of the recombinant gene. Four antimicrobial peptides with strong bactericidal properties served as toxic gene products, while green fluorescent protein was used to assess the efficiency of expression repression. As a result, we developed an expression system with strong repression, which is activated by CRISPR-Cas9-mediated excision of a DNA fragment from the plasmids.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2025-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144638679","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Implementation of an Undergraduate Laboratory-Based Mammalian Genome Editing Course. 基于实验室的本科哺乳动物基因组编辑课程的实施。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2025-07-10 DOI: 10.1089/crispr.2025.0017

Isabelle Guerra, Karin Jensen, Pablo Perez-Pinera

{"title":"Implementation of an Undergraduate Laboratory-Based Mammalian Genome Editing Course.","authors":"Isabelle Guerra, Karin Jensen, Pablo Perez-Pinera","doi":"10.1089/crispr.2025.0017","DOIUrl":"https://doi.org/10.1089/crispr.2025.0017","url":null,"abstract":"Genome engineering methods can be utilized to perform complex genetic manipulations in living cells with remarkable efficiency and precision. Given the transformative potential of these enabling technologies, their applications are steadily expanding into most biology and biomedical fields where they play a central role in many experimental frameworks. For these reasons, in order to effectively prepare future generations of biologists and bioengineers for successful careers, there is a high need to incorporate courses teaching genome editing fundamentals into existing curricula. To accomplish this objective, lecture-based courses are rapidly integrating genome editing concepts; however, there are few laboratory courses that teach the practical skills needed to successfully perform genome editing experiments. Here, we describe the development and implementation of a semester-long laboratory course that teaches students not only the techniques needed to perform gene knockout, gene activation, gene repression, and base editing in mammalian cells but also prepares them to design and troubleshoot experiments, write scientific manuscripts, as well as prepare and deliver scientific presentations. Course evaluations demonstrate that this class effectively equips students with the knowledge and hands-on experience needed to succeed in careers related to genome engineering, cell and tissue engineering, and, more broadly, biology.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144602242","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Advances in Gene Therapy for Sickle Cell Disease: From Preclinical Innovations to Clinical Implementation and Access Challenges. 镰状细胞病基因治疗的进展：从临床前创新到临床实施和获取挑战。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2025-06-01 Epub Date: 2025-05-12 DOI: 10.1089/crispr.2024.0101

Henna Butt, Mamatha Mandava, David Jacobsohn

引用次数: 0

CRISPR2025 New Zealand: Innovation and Collaboration. CRISPR2025新西兰：创新与合作。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2025-06-01 Epub Date: 2025-05-29 DOI: 10.1089/crispr.2025.0026

Katharina G Wandera, Jeremy Dubrulle, Russell Greene, Meric Ozturk, Gavin Knott, Dipali G Sashital, Peter C Fineran

引用次数: 0

Metagenome-Derived CRISPR-Cas12a Mining and Characterization. 宏基因组衍生的CRISPR-Cas12a挖掘和表征。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2025-06-01 Epub Date: 2025-05-21 DOI: 10.1089/crispr.2024.0099

Kalani Gast, Sydney Baker, Adair L Borges, Stephanie Ward, Jillian F Banfield, Rodolphe Barrangou

{"title":"Metagenome-Derived CRISPR-Cas12a Mining and Characterization.","authors":"Kalani Gast, Sydney Baker, Adair L Borges, Stephanie Ward, Jillian F Banfield, Rodolphe Barrangou","doi":"10.1089/crispr.2024.0099","DOIUrl":"10.1089/crispr.2024.0099","url":null,"abstract":"The advent of clustered regularly interspaced short palindromic repeats (CRISPR)-based technologies has revolutionized genome editing, with continued interest in expanding the CRISPR-associated proteins (Cas) toolbox with diverse, efficient, and specific effectors. CRISPR-Cas12a is a potent, programmable RNA-guided dual nickase, broadly used for genome editing. Here, we mined dairy cow microbial metagenomes for CRISPR-Cas systems, unraveling novel Cas12a enzymes. Using in silico pipelines, we characterized and predicted key drivers of CRISPR-Cas12a activity, encompassing guides and protospacer adjacent motifs for five systems. We next assessed their functional potential in cell-free transcription-translation assays with GFP-based fluorescence readouts. Lastly, we determined their genome editing potential in vivo in Escherichia coli by generating 1 kb knockouts. Unexpectedly, we observed natural sequence variation in the bridge-helix domain of the best-performing candidate and used mutagenesis to alter the activity of Cas12a orthologs, resulting in increased gene editing capabilities of a relatively inefficient candidate. This study illustrates the potential of underexplored metagenomic sequence diversity for the development and refinement of genome editing effectors.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"189-204"},"PeriodicalIF":3.7,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144121294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Facing Systemic Uncertainty, Can the CRISPR Community Stay the Course? 面对系统的不确定性，CRISPR社区能坚持到底吗？

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2025-06-01 DOI: 10.1089/crispr.2025.0050

Rodolphe Barrangou

引用次数: 0

Green Listed v2.0: A Web Application for Streamlined Design of Custom CRISPR Screens. 绿色列表v2.0：自定义CRISPR屏幕的流线型设计的Web应用程序。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2025-06-01 Epub Date: 2025-05-07 DOI: 10.1089/crispr.2025.0023

Esbjörn Henkel, Zhaojun Li, Daniel Uvehag, Bernhard Schmierer, Martin Henkel, Fredrik Wermeling

引用次数: 0

Optimizing Genome Editing in Mollusks (Crassostrea gigas) in Vitro Validation of sgRNA and Identifying Key Factors Influencing Efficiency. 优化软体动物（长牡蛎）基因组编辑体外验证sgRNA及确定影响效率的关键因素

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2025-06-01 Epub Date: 2025-02-28 DOI: 10.1089/crispr.2024.0086

Qian Li, Hong Yu, Shaojun Du, Qi Li

{"title":"Optimizing Genome Editing in Mollusks (Crassostrea gigas) in Vitro Validation of sgRNA and Identifying Key Factors Influencing Efficiency.","authors":"Qian Li, Hong Yu, Shaojun Du, Qi Li","doi":"10.1089/crispr.2024.0086","DOIUrl":"10.1089/crispr.2024.0086","url":null,"abstract":"CRISPR-Cas9 genome editing holds tremendous potential for accelerating genetic improvements in aquaculture. The success of the CRISPR-Cas9 system relies on the specificity and efficiency of engineered single-guide RNAs (sgRNAs). In this study, we optimized an in vitro validation protocol for sgRNAs to streamline the gene editing process, capitalizing on the limited breeding season of the Pacific oyster (Crassostrea gigas). We evaluated the efficiency of 11 sgRNAs targeting four genes both in vitro and in vivo in C. gigas. In addition, we found that Cas9 protein differs from Cas9 mRNA in gene editing efficiency at various stages of early development. Cas9 protein proved particular efficacy in achieving early and efficient gene knockout, functioning effectively during the first cell division and facilitating biallelic gene knockouts. Statistical analysis showed that in the protein group, the biallelic editing frequency ranged from 12.5% to 57.8%, and the overall editing frequency reached as high as 75-90.6%. The mRNA group exhibited a biallelic editing frequency of 3.1-14.0% and the overall editing frequency spanning 65.6-78.1%. Contrary to expectations, low-temperature incubation (20°C) of oyster embryos prolonged the time window for the first cell division but did not improve gene editing efficiency, likely due to the high temperature sensitivity of Cas9 enzyme activity. Together, this study provides a comprehensive analysis of factors affecting the efficiency of CRISPR-Cas9 gene editing in C. gigas, providing a robust framework for future gene editing endeavors in mollusks and other marine invertebrates.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"205-215"},"PeriodicalIF":3.7,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143528008","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Biotechnologies in the World: On Global Asymmetries and the Need for Cosmopolitanism. 世界上的生物技术：关于全球不对称和世界主义的需要。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2025-05-21 DOI: 10.1089/crispr.2025.0054

Kaushik Sunder Rajan

引用次数: 0