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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 4 4区 生物学
CRISPR Journal Pub Date : 2025-10-01 Epub 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":"10.1177/25731599251358852","url":null,"abstract":"<p><p>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 <i>Escherichia coli</i> 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.</p>","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"353-365"},"PeriodicalIF":4.0,"publicationDate":"2025-10-01","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
A Phage Variable Region Encodes Anti-CRISPR Proteins Inhibiting All Streptococcus thermophilus CRISPR Immune Systems. 噬菌体可变区编码抗CRISPR蛋白抑制所有嗜热链球菌CRISPR免疫系统。
IF 4 4区 生物学
CRISPR Journal Pub Date : 2025-10-01 Epub Date: 2025-09-29 DOI: 10.1177/25731599251369720
Katie A Johnson, Clare Cooper, Cécile Philippe, Ryan J Catchpole, Shakela Mitchell, Michael P Terns
{"title":"A Phage Variable Region Encodes Anti-CRISPR Proteins Inhibiting All <i>Streptococcus thermophilus</i> CRISPR Immune Systems.","authors":"Katie A Johnson, Clare Cooper, Cécile Philippe, Ryan J Catchpole, Shakela Mitchell, Michael P Terns","doi":"10.1177/25731599251369720","DOIUrl":"10.1177/25731599251369720","url":null,"abstract":"<p><p>Bacteria and archaea utilize CRISPR-Cas systems to defend against invading mobile genetic elements (MGEs) such as phages and plasmids. In turn, MGEs have evolved anti-CRISPR (Acr) proteins to counteract these defenses. While several type II-A Acrs have been identified in <i>Streptococcus thermophilus</i> (<i>Sth</i>) phages, a more comprehensive understanding of Acr diversity in <i>Sth</i> phages has yet to be explored. Guided by the genomic context of known Acrs, we systematically screened uncharacterized phage proteins and identified several novel Acrs that inhibit type I-E, type II-A or type III-A <i>Sth</i> CRISPR-Cas systems. These <i>acr</i> genes are clustered within a variable phage genomic region, indicating a hotspot for anti-defense activity. We also identified neighboring proteins with predicted enzymatic or structural domains that may modulate phage-host interactions through Acr-independent mechanisms. Together, our findings expand the known repertoire of <i>Sth</i> Acrs and highlight the phage variable region as a key reservoir of immune-modulating factors.</p>","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"333-352"},"PeriodicalIF":4.0,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145187423","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 4 4区 生物学
CRISPR Journal Pub Date : 2025-10-01 Epub 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":"10.1089/crispr.2025.0017","url":null,"abstract":"<p><p>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.</p>","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"366-374"},"PeriodicalIF":4.0,"publicationDate":"2025-10-01","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
Efficient CRISPR-Cas9-Mediated Genome Editing of the Cane Toad (Rhinella marina). 高效crispr - cas9介导的甘蔗蟾蜍基因组编辑
IF 4 4区 生物学
CRISPR Journal Pub Date : 2025-10-01 Epub Date: 2025-09-26 DOI: 10.1177/25731599251382427
Michael B Clark, Alexander T Funk, Alex Paporakis, Gregory P Brown, Samuel J Beach, Aidan Tay, Stephanie Deering, Caitlin Cooper, Mark Tizard, Chris J Jolly, Georgia Ward-Fear, Anthony W Waddle, Richard Shine, Maciej Maselko
{"title":"Efficient CRISPR-Cas9-Mediated Genome Editing of the Cane Toad (<i>Rhinella marina</i>).","authors":"Michael B Clark, Alexander T Funk, Alex Paporakis, Gregory P Brown, Samuel J Beach, Aidan Tay, Stephanie Deering, Caitlin Cooper, Mark Tizard, Chris J Jolly, Georgia Ward-Fear, Anthony W Waddle, Richard Shine, Maciej Maselko","doi":"10.1177/25731599251382427","DOIUrl":"10.1177/25731599251382427","url":null,"abstract":"<p><p>Invasive species inflict major ecological, economic, and cultural harm worldwide, highlighting the urgent need for innovative control strategies. Genome editing offers exciting possibilities for targeted control methods for invasive species. Here, we demonstrate CRISPR-Cas9 genome editing in the cane toad (<i>Rhinella marina</i>), one of Australia's most notorious invasive species, by targeting the <i>tyrosinase</i> gene to produce albino phenotypes as visual markers for assessing editing efficiency. Microinjection of Cas9 protein and guide RNAs into one-cell zygotes resulted in 87.6% of mosaic larvae displaying nearly complete albinism, with 2.3% exhibiting complete albinism. For completely albino individuals, genomic analysis confirmed predominantly frameshift mutations or large deletions at the target site, with no wild-type alleles detected. Germline transmission rates reflected the extent of albinism in the mosaic adult, with maternal transmission approaching 100%. This first application of CRISPR-Cas9 in the Bufonidae family opens possibilities for exploring basic research questions and population control strategies.</p>","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"321-332"},"PeriodicalIF":4.0,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145151918","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 4 4区 生物学
CRISPR Journal Pub Date : 2025-08-01 Epub Date: 2025-05-21 DOI: 10.1089/crispr.2025.0054
Kaushik Sunder Rajan
{"title":"Biotechnologies in the World: On Global Asymmetries and the Need for Cosmopolitanism.","authors":"Kaushik Sunder Rajan","doi":"10.1089/crispr.2025.0054","DOIUrl":"10.1089/crispr.2025.0054","url":null,"abstract":"<p><p>Conversations regarding genome editing are not simply about the transformative science involved. They touch upon fundamental moral questions concerning the human condition, indeed what it means to be human itself. The recent approval of a gene therapy for sickle cell disease encapsulates the relationship between scientific innovation and health care access and the relations of power and political economy that structure the world of biotech and biomedicine. Globally transformative biotechnologies must ethically situate themselves if they are not merely to reproduce longstanding historical and structural asymmetries. The time has come to embrace a cosmopolitan ethic that is attuned to the varied constitutionalisms through which debates about public good, healthy societies, and social compacts materialize around the world.</p>","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"288-292"},"PeriodicalIF":4.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144112739","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
Science with Society at Science Gallery Bengaluru. 科学与社会在班加罗尔科学画廊。
IF 4 4区 生物学
CRISPR Journal Pub Date : 2025-08-01 Epub Date: 2025-05-20 DOI: 10.1089/crispr.2025.0052
Jahnavi Phalkey
{"title":"Science with Society at Science Gallery Bengaluru.","authors":"Jahnavi Phalkey","doi":"10.1089/crispr.2025.0052","DOIUrl":"10.1089/crispr.2025.0052","url":null,"abstract":"<p><p>Science Gallery Bengaluru was established to serve as a two-way bridge between the public and new and old research. This work is furthered through public engagement in the form of year-long living exhibitions, a public laboratory complex with five experimental spaces, and a mentorship initiative for young adults. There is a strong conversation about professions in science and engineering in the Indian public domain, but its cultural equivalent is less developed. In this context, ideas and topics for exploration at the Gallery are chosen not for novelty with which the public is to be familiarized, but for their already robust presence in public discourse in order to explore their complexity through research in the human, social, and natural sciences, with a view to enabling visitors to make more informed choices in everyday life.</p>","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"282-284"},"PeriodicalIF":4.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144112751","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
Summitting CRISPR for Human Heritable Genome Editing. 人类可遗传基因组编辑CRISPR峰会。
IF 4 4区 生物学
CRISPR Journal Pub Date : 2025-08-01 Epub Date: 2025-05-21 DOI: 10.1089/crispr.2025.0051
Françoise Baylis
{"title":"Summitting CRISPR for Human Heritable Genome Editing.","authors":"Françoise Baylis","doi":"10.1089/crispr.2025.0051","DOIUrl":"10.1089/crispr.2025.0051","url":null,"abstract":"<p><p>The ethical issues of human heritable genome editing have been discussed at international summits held since 2015. In this Perspective, I consider how the discussions evolved over three summits held in Washington, DC (2015), Hong Kong (2018), and London (2023). The significance of safety and efficacy, meanings of a moratorium, and place of broad societal consensus are traced through publications produced surrounding these summits. Looking ahead, I highlight the difference between two fundamentally distinct ethical questions: Is human heritable genome editing ethical? Can human heritable genome editing be done ethically?</p>","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"239-244"},"PeriodicalIF":4.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144112753","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
Introducing Faith-Based Claims to Ethical Discourse on Scientific Matters. 将基于信仰的主张引入科学问题的伦理论述。
IF 4 4区 生物学
CRISPR Journal Pub Date : 2025-08-01 Epub Date: 2025-05-21 DOI: 10.1089/crispr.2025.0045
James F Keenan
{"title":"Introducing Faith-Based Claims to Ethical Discourse on Scientific Matters.","authors":"James F Keenan","doi":"10.1089/crispr.2025.0045","DOIUrl":"10.1089/crispr.2025.0045","url":null,"abstract":"<p><p>For the majority of the human community, the questions posed by heritable genome editing regarding the meanings of human life draw on various religious perspectives. Deliberations about genome editing would benefit from thinking about religious concerns in non-reductionist and non-instrumental terms. While it is often suggested that the scientific community is intolerant of religious views, it is often not the scientific community but those working in philosophy and religion who self-censor religious language. Rather than merely including people who are conversant in the language of religion or trying to derive an ethical Esperanto from generic religious beliefs, I advocate in this Perspective for the inclusion of people who voice their ethical commitments in the theological principles of their particular religious commitments.</p>","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"307-310"},"PeriodicalIF":4.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144112746","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
Moving Therapeutic Genome Editing into Global Clinical Trials and Medicine. 将治疗性基因组编辑推向全球临床试验和医学。
IF 4 4区 生物学
CRISPR Journal Pub Date : 2025-08-01 Epub Date: 2025-05-22 DOI: 10.1089/crispr.2025.0049
Kiran Musunuru, Fyodor Urnov
{"title":"Moving Therapeutic Genome Editing into Global Clinical Trials and Medicine.","authors":"Kiran Musunuru, Fyodor Urnov","doi":"10.1089/crispr.2025.0049","DOIUrl":"10.1089/crispr.2025.0049","url":null,"abstract":"<p><p>Moving CRISPR-based therapies from discovery to dosing patients in clinical trials and ultimately to approval involves navigating a challenging terrain of highs and lows. In this interview, physician-scientist Kiran Musunuru and genome editor Fyodor Urnov reflect on the past 20 years of their nonclinical and clinical programs in the field, the current landscape of innovation, and what they see on the horizon.</p>","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"228-231"},"PeriodicalIF":4.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144112748","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
Intent to Cure: The Need for a Rare Disease Platform in India and Across the Global South. 治愈的意图：印度和全球南方需要一个罕见疾病平台。
IF 4 4区 生物学
CRISPR Journal Pub Date : 2025-08-01 Epub Date: 2025-05-21 DOI: 10.1089/crispr.2025.0046
Vijay Chandru, Vaijayanti Gupta, Vandana Hegde, Arvind Venkatesan, Reety Arora
{"title":"Intent to Cure: The Need for a Rare Disease Platform in India and Across the Global South.","authors":"Vijay Chandru, Vaijayanti Gupta, Vandana Hegde, Arvind Venkatesan, Reety Arora","doi":"10.1089/crispr.2025.0046","DOIUrl":"10.1089/crispr.2025.0046","url":null,"abstract":"<p><p>The democratization of genomic technologies presents substantial opportunities for addressing rare genetic diseases, particularly in collaborations between the Global South and North. In this Perspective, we describe the current progress in gene therapy, including CRISPR, in India and see an upward trajectory of innovation. We propose the establishment of a rare disease platform in India and across the Global South designed to bridge scientific, clinical, and economic gaps, transforming untapped genetic diversity into shared opportunities for therapeutic innovation and health care equity. This platform would encompass a comprehensive data infrastructure capturing clinical, genomic, and biosample data, complemented by an artificial intelligence-powered analytics layer to enhance patient engagement and clinical trial matching, ultimately enabling cost-effective research and development (R&D) of novel therapies.</p>","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"277-281"},"PeriodicalIF":4.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144112745","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
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