CRISPR Journal最新文献_第5页

Novel off-Targeting Events Identified after Genome Wide Analysis of CRISPR-Cas Edited Pigs. 对 CRISPR-Cas 编辑过的猪进行全基因组分析后发现的新的非靶向事件。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-06-01 Epub Date: 2024-05-21 DOI: 10.1089/crispr.2024.0012

Bethany K Redel, Junchul Yoon, Emily Reese, Hong An, Kyungjun Uh, Paula R Chen, Randall S Prather, Kiho Lee

{"title":"Novel off-Targeting Events Identified after Genome Wide Analysis of CRISPR-Cas Edited Pigs.","authors":"Bethany K Redel, Junchul Yoon, Emily Reese, Hong An, Kyungjun Uh, Paula R Chen, Randall S Prather, Kiho Lee","doi":"10.1089/crispr.2024.0012","DOIUrl":"10.1089/crispr.2024.0012","url":null,"abstract":"CRISPR-Cas technology has transformed our ability to introduce targeted modifications, allowing unconventional animal models such as pigs to model human diseases and improve its value for food production. The main concern with using the technology is the possibility of introducing unwanted modifications in the genome. In this study, we illustrate a pipeline to comprehensively identify off-targeting events on a global scale in the genome of three different gene-edited pig models. Whole genome sequencing paired with an off-targeting prediction software tool filtered off-targeting events amongst natural variations present in gene-edited pigs. This pipeline confirmed two known off-targeting events in IGH knockout pigs, AR and RBFOX1, and identified other presumably off-targeted loci. Independent validation of the off-targeting events using other gene-edited DNA confirmed two novel off-targeting events in RAG2/IL2RG knockout pig models. This unique strategy offers a novel tool to detect off-targeting events in genetically heterogeneous species after genome editing.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"141-149"},"PeriodicalIF":3.7,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11302191/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141072230","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A Story of Perseverance: An Interview with Matthew Porteus. 坚持不懈的故事：马修-波特斯访谈录

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-06-01 DOI: 10.1089/crispr.2024.0043

Matthew H Porteus, Kevin Davies

引用次数: 0

Surveying the State of CRISPR and Gene Editing. 调查 CRISPR 和基因编辑的现状。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-06-01 DOI: 10.1089/crispr.2024.0045

Rodolphe Barrangou

引用次数: 0

Establishment of a Cleavage-Based Single-Plasmid Dual-Luciferase Surrogate Reporter for the Cleavage Efficiency Evaluation of CRISPR-Cas12a Systems and Its Primary Application. 建立基于裂解的单质粒双荧光素酶替代报告物，用于评估 CRISPR-Cas12a 系统的裂解效率及其主要应用。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-06-01 DOI: 10.1089/crispr.2024.0038

Yaoqiang Shi, Qi Tan, Chunhui Yang, Shilin Li, Yujia Li, Baoren He, He Xie, Xiaoqiong Duan, Limin Chen

{"title":"Establishment of a Cleavage-Based Single-Plasmid Dual-Luciferase Surrogate Reporter for the Cleavage Efficiency Evaluation of CRISPR-Cas12a Systems and Its Primary Application.","authors":"Yaoqiang Shi, Qi Tan, Chunhui Yang, Shilin Li, Yujia Li, Baoren He, He Xie, Xiaoqiong Duan, Limin Chen","doi":"10.1089/crispr.2024.0038","DOIUrl":"https://doi.org/10.1089/crispr.2024.0038","url":null,"abstract":"CRISPR-Cas technology is a widely utilized gene-editing tool that involves gRNA-guided sequence recognition and Cas nuclease-mediated cleavage. The design and evaluation of gRNA are essential for enhancing CRISPR/Cas editing efficiency. Various assays such as single-strand annealing, in vitro cleavage, and T7 endonuclease I (T7EI) are commonly used to assess gRNA-mediated Cas protein cleavage activity. In this study, a firefly luciferase and Renilla luciferase co-expressed and a cleavage-based single-plasmid dual-luciferase surrogate reporter was built to evaluate the gRNA-mediated Cas12a cleavage efficiency. The cleavage activities of CRISPR-Cas12a can be quantitatively determined by the recovery degree of firefly luciferase activity. The cleavage efficiency of CRISPR-Cas12a can be quantitatively measured by the recovery of firefly luciferase activity. By using this system, the cleavage efficiency of CRISPR-Cas12a on hepatitis B virus (HBV)/D expression plasmid was evaluated, revealing a negative correlation between gRNA cleavage efficiency and HBV gene expression measured using an enzyme-linked immunosorbent assay. This simple, efficient, and quantifiable system only requires the dual-luciferase vector and CRISPR-Cas12a vector, making it a valuable tool for selecting effective gRNAs for gene editing.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":"7 3","pages":"156-167"},"PeriodicalIF":3.7,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141452152","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

Making the Search for Genome Editing Tortured Phrases a Collective Effort. 让寻找基因组编辑折磨人的短语成为一项集体工作。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-04-01 DOI: 10.1089/crispr.2024.0015

Guillaume Levrier

引用次数: 0

Gene Editing in the Chagas Disease Vector Rhodnius prolixus by Cas9-Mediated ReMOT Control. 通过 Cas9 介导的 ReMOT 控制对南美锥虫病病媒 Rhodnius prolixus 进行基因编辑。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-04-01 DOI: 10.1089/crispr.2023.0076

Leonardo Lima, Mateus Berni, Jamile Mota, Daniel Bressan, Alison Julio, Robson Cavalcante, Vanessa Macias, Zhiqian Li, Jason L Rasgon, Ethan Bier, Helena Araujo

{"title":"Gene Editing in the Chagas Disease Vector Rhodnius prolixus by Cas9-Mediated ReMOT Control.","authors":"Leonardo Lima, Mateus Berni, Jamile Mota, Daniel Bressan, Alison Julio, Robson Cavalcante, Vanessa Macias, Zhiqian Li, Jason L Rasgon, Ethan Bier, Helena Araujo","doi":"10.1089/crispr.2023.0076","DOIUrl":"10.1089/crispr.2023.0076","url":null,"abstract":"Rhodnius prolixus is currently the model vector of choice for studying Chagas disease transmission, a debilitating disease caused by Trypanosoma cruzi parasites. However, transgenesis and gene editing protocols to advance the field are still lacking. Here, we tested protocols for the maternal delivery of CRISPR-Cas9 (clustered regularly spaced palindromic repeats/Cas-9 associated) elements to developing R. prolixus oocytes and strategies for the identification of insertions and deletions (indels) in target loci of resulting gene-edited generation zero (G0) nymphs. We demonstrate successful gene editing of the eye color markers Rp-scarlet and Rp-white, and the cuticle color marker Rp-yellow, with highest effectiveness obtained using Receptor-Mediated Ovary Transduction of Cargo (ReMOT Control) with the ovary-targeting BtKV ligand. These results provide proof of concepts for generating somatic mutations in R. prolixus and potentially for generating germ line-edited lines in triatomines, laying the foundation for gene editing protocols that could lead to the development of novel control strategies for vectors of Chagas disease.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"88-99"},"PeriodicalIF":3.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140337648","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

Acknowledgment of Reviewers 2023. 鸣谢 2023 年审稿人。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-02-01 DOI: 10.1089/crispr.2024.29169.ack

引用次数: 0

Correction to: Programmable RNA Targeting Using CasRx in Flies, by Buchman, et al. The CRISPR Journal 2020;3(3)164-176; doi: 10.1089/crispr.2020.0018. 更正：Buchman 等人撰写的《利用 CasRx 在蝇类中进行可编程 RNA 靶向》（Programmable RNA Targeting Using CasRx in Flies）。 CRISPR 期刊》2020; 3(3)164-176; doi: 10.1089/crispr.2020.0018。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-02-01 DOI: 10.1089/crispr.2020.0018.correx

引用次数: 0

CRISPR Momentum in the Clinic and the Field. CRISPR 在临床和科研领域的发展势头。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-02-01 DOI: 10.1089/crispr.2024.29172.editorial