CRISPR Journal最新文献_第3页

Repurposing an Endogenous CRISPR-Cas System to Generate and Study Subtle Mutations in Bacteriophages. 重新利用内源性 CRISPR-Cas 系统，生成并研究噬菌体中的微妙突变。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-12-01 Epub Date: 2024-09-30 DOI: 10.1089/crispr.2024.0047

Kotaro Kamata, Nils Birkholz, Marijn Ceelen, Robert D Fagerlund, Simon A Jackson, Peter C Fineran

{"title":"Repurposing an Endogenous CRISPR-Cas System to Generate and Study Subtle Mutations in Bacteriophages.","authors":"Kotaro Kamata, Nils Birkholz, Marijn Ceelen, Robert D Fagerlund, Simon A Jackson, Peter C Fineran","doi":"10.1089/crispr.2024.0047","DOIUrl":"10.1089/crispr.2024.0047","url":null,"abstract":"While bacteriophage applications benefit from effective phage engineering, selecting the desired genotype after subtle modifications remains challenging. Here, we describe a two-phase endogenous CRISPR-Cas-based phage engineering approach that enables selection of small defined edits in Pectobacterium carotovorum phage ZF40. We designed plasmids containing sequences homologous to ZF40 and a mini-CRISPR array. The plasmids allowed genome editing through homologous recombination and counter-selection against non-recombinant phage genomes using an endogenous type I-E CRISPR-Cas system. With this technique, we first deleted target genes and subsequently restored loci with modifications. This two-phase approach circumvented major challenges in subtle phage modifications, including inadequate sequence distinction for CRISPR-Cas counter-selection and the requirement of a protospacer-adjacent motif, limiting sequences that can be modified. Distinct 20-bp barcodes were incorporated through engineering as differential target sites for programmed CRISPR-Cas activity, which allowed quantification of phage variants in mixed populations. This method aids studies and applications that require mixtures of similar phages.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"343-354"},"PeriodicalIF":3.7,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142332259","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

Mutation-Specific CRISPR Targeting with SaCas9 and AsCas12a Restores Therapeutic Sensitivity in Treatment-Resistant Melanoma. 用 SaCas9 和 AsCas12a 进行突变特异性 CRISPR 靶向可恢复耐药黑色素瘤的治疗敏感性。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-12-01 Epub Date: 2024-10-10 DOI: 10.1089/crispr.2024.0003

Brett M Sansbury, Sophia B Masciarelli, Salma Kaouser, Olivia M Tharp, Kelly H Banas, Eric B Kmiec

{"title":"Mutation-Specific CRISPR Targeting with SaCas9 and AsCas12a Restores Therapeutic Sensitivity in Treatment-Resistant Melanoma.","authors":"Brett M Sansbury, Sophia B Masciarelli, Salma Kaouser, Olivia M Tharp, Kelly H Banas, Eric B Kmiec","doi":"10.1089/crispr.2024.0003","DOIUrl":"10.1089/crispr.2024.0003","url":null,"abstract":"Background: Melanoma remains one of the most challenging cancers to treat effectively with drug resistant remaining a constant concern, primarily with activating BRAF mutations. Mutations in the BRAF gene appear in approximately 50% of patients, 90% of which are V600E. Two frontline BRAF inhibitors (BRAFi), vemurafenib and dabrafenib, are frequently used to treat unresectable or metastatic BRAF V600E melanoma. Initial response rates are high, but soon thereafter, 70-80% of patients develop resistance to treatment within a year. A major mechanism of resistance is the generation of a secondary Q61K mutation in the NRAS gene. Methods: We have developed an approach in which a CRISPR-Cas complex can be designed to distinguish between mutant genes enabling resistance to standard care in tumor cells and normal genomes of healthy cells. For the first time, we demonstrated the utility of two CRISPR-directed mutation-specific editing approaches to restore BRAFi sensitivity in BRAFV600E/NRASQ61K resistant A375 cells. Results: We utilize an AsCas12a protospacer adjacent motif site created by the NRAS Q61K mutation and the Q61K mutation in the critical seed region of an SaCas9 sgRNA for Q61K-selective targeting. We show here that both approaches allow for effective NRAS targeting of only mutated-Q61K and after CRISPR-directed Q61K-targeting, previously resistant A375 cells are re-sensitized to BRAFi treatment. Conclusion: Our data support the feasibility of the development of CRISPR-Cas therapeutic approaches to the treatment of melanoma. Successful therapeutic CRISPR-directed gene editing would enable both specific and efficient editing of a mutation-specific targeting approach eliminate concern for on- and off-target damage to the genomes of healthy cells.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"366-373"},"PeriodicalIF":3.7,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142480624","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

Engineering of HEK293T Cell Factory for Lentiviral Production by High-Throughput Selected Genes. 通过高通量选择基因对 HEK293T 细胞工厂进行工程改造，以生产慢病毒。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-10-01 Epub Date: 2024-10-16 DOI: 10.1089/crispr.2024.0016

Zhang Xinyue, Siwei Li, Wang Yujie, Dai Yingcai, Bi Changhao, Zhang Xueli

{"title":"Engineering of HEK293T Cell Factory for Lentiviral Production by High-Throughput Selected Genes.","authors":"Zhang Xinyue, Siwei Li, Wang Yujie, Dai Yingcai, Bi Changhao, Zhang Xueli","doi":"10.1089/crispr.2024.0016","DOIUrl":"10.1089/crispr.2024.0016","url":null,"abstract":"Lentiviral vectors (LVs) are crucial tools in gene therapy and bioproduction, but high-yield LV production systems are urgently needed. Using clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 high-throughput screening, we identified nine critical genes (LDAH, GBP3, BPIFC, NHLRC1, NHLRC3, ZNF425, TTC37, LRRC4B, and SPINK6) from 17,501 genes that limit LV packaging and formation. Knocking out these genes in HEK293T cells significantly increased virus production, with LDAH knockout exhibiting a 6.63-fold increase. Studies on multigene knockouts demonstrated that the cumulative effects of different gene knockouts can significantly enhance lentivirus production in HEK293T cells. Triple knockout of GBP3, BPIFC, and LDAH increased LV titer by ∼8.33-fold, and knockout (or knockdown) of GBP3, NHLRC1, and NHLRC3 increased LV titer by ∼6.53-fold. This study established HEK293T cell lines with multiple genes knockout for efficient LV production, providing reliable technical support for LV production and application and offering new perspectives for studying LV packaging mechanisms and related virus research.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":"7 5","pages":"272-282"},"PeriodicalIF":3.7,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142480626","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

Rosalind Franklin Society Proudly Announces the 2023 Award Recipient for The CRISPR Journal. 罗莎琳德·富兰克林协会自豪地宣布了2023年CRISPR杂志的获奖者。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-10-01 DOI: 10.1089/crispr.2024.55675.rfs2023

Suchita Nety

引用次数: 0

Five Years of Progress in CRISPR Clinical Trials (2019-2024). CRISPR 临床试验的五年进展（2019-2024 年）。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-10-01 DOI: 10.1089/crispr.2024.0081

Kevin Davies, Alex Philippidis, Rodolphe Barrangou

引用次数: 0

Characterization of Research Support of Genome Editing Technologies and Transition to Clinical Trials. 基因组编辑技术研究支持的特点以及向临床试验的过渡。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-10-01 Epub Date: 2024-09-26 DOI: 10.1089/crispr.2024.0011

Riya Mohan, Susanne B Haga

引用次数: 0

Genome Editing Therapy for the Blood: Ex Vivo Success and In Vivo Prospects. 血液基因组编辑疗法：体内成功与体内前景。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-10-01 Epub Date: 2024-09-26 DOI: 10.1089/crispr.2024.0036

Christy A George, Srishti U Sahu, Lorena de Oñate, Bruno Solano de Freitas Souza, Ross C Wilson

{"title":"Genome Editing Therapy for the Blood: Ex Vivo Success and In Vivo Prospects.","authors":"Christy A George, Srishti U Sahu, Lorena de Oñate, Bruno Solano de Freitas Souza, Ross C Wilson","doi":"10.1089/crispr.2024.0036","DOIUrl":"10.1089/crispr.2024.0036","url":null,"abstract":"Hematopoietic stem cells (HSCs) provide the body with a continuous supply of healthy, functional blood cells. In patients with hematopoietic malignancies, immunodeficiencies, lysosomal storage disorders, and hemoglobinopathies, therapeutic genome editing offers hope for corrective intervention, with even modest editing efficiencies likely to provide clinical benefit. Engineered white blood cells, such as T cells, can be applied therapeutically to address monogenic disorders of the immune system, HIV infection, or cancer. The versatility of CRISPR-based tools allows countless new medical interventions for diseases of the blood, and rapid ex vivo success has been demonstrated in hemoglobinopathies via transplantation of the patient's HSCs following genome editing in a laboratory setting. Here we review recent advances in therapeutic genome editing of HSCs and T cells, focusing on the progress in ex vivo contexts, the promise of improved access via in vivo delivery, as well as the ongoing preclinical efforts that may enable the transition from ex vivo to in vivo administration. We discuss the challenges, limitations, and future prospects of this rapidly developing field, which may one day establish CRISPR as the standard of care for some diseases affecting the blood.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"231-248"},"PeriodicalIF":3.7,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142332258","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

Rosalind Franklin Society Proudly Announces the 2023 Award Recipient for The CRISPR Journal. 罗莎琳德-富兰克林学会自豪地宣布《CRISPR 期刊》2023 年度获奖者。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-10-01 DOI: 10.1089/crispr.2024.55675.rfs2023

Suchita Nety

引用次数: 0

Comparison of Gene-Editing Approaches for Severe Congenital Neutropenia-Causing Mutations in the ELANE Gene. 比较针对 ELANE 基因中导致严重先天性中性粒细胞减少症突变的基因编辑方法

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-10-01 DOI: 10.1089/crispr.2024.0006

Malte Ulrich Ritter, Masoud Nasri, Benjamin Dannenmann, Perihan Mir, Benjamin Secker, Diana Amend, Maksim Klimiankou, Karl Welte, Julia Skokowa

{"title":"Comparison of Gene-Editing Approaches for Severe Congenital Neutropenia-Causing Mutations in the ELANE Gene.","authors":"Malte Ulrich Ritter, Masoud Nasri, Benjamin Dannenmann, Perihan Mir, Benjamin Secker, Diana Amend, Maksim Klimiankou, Karl Welte, Julia Skokowa","doi":"10.1089/crispr.2024.0006","DOIUrl":"10.1089/crispr.2024.0006","url":null,"abstract":"Safety considerations for gene therapies of inherited preleukemia syndromes, including severe congenital neutropenia (CN), are paramount. We compared several strategies for CRISPR/Cas9 gene editing of autosomal-dominant ELANE mutations in CD34+ cells from two CN patients head-to-head. We tested universal and allele-specific ELANE knockout, ELANE mutation correction by homology-directed repair (HDR) with AAV6, and allele-specific HDR with ssODN. All strategies were not toxic, had at least 30% editing, and rescued granulopoiesis in vitro. In contrast to published data, allele-specific indels in the last exon of ELANE also restored granulopoiesis. Moreover, by implementing patient-derived induced pluripotent stem cells for GUIDE-Seq off-target analysis, we established a clinically relevant \"personalized\" assessment of off-target activity of gene editing on the background of the patient's genome. We found that allele-specific approaches had the most favorable off-target profiles. Taken together, a well-defined head-to-head comparison pipeline for selecting the appropriate gene therapy is essential for diseases, with several gene editing strategies available.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":"7 5","pages":"258-271"},"PeriodicalIF":3.7,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142480625","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

Give Cas a Chance: An Actionable Path to a Platform for CRISPR Cures. 给 Cas 一个机会：通往 CRISPR 治疗平台的可行之路。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-10-01 DOI: 10.1089/crispr.2024.0082

Fyodor D Urnov

引用次数: 0