CRISPR Journal最新文献_第9页

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

Rodolphe Barrangou, Kevin Davies

引用次数: 0

Warrior Spirit: An Interview with Victoria Gray, Sickle Cell Pioneer. 勇士精神：镰状细胞先驱维多利亚-格雷访谈录。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-02-01 DOI: 10.1089/crispr.2024.29171.vgr

Victoria Gray, Uduak Thomas, Kevin Davies

引用次数: 0

Generation of a Commercial-Scale Founder Population of Porcine Reproductive and Respiratory Syndrome Virus Resistant Pigs Using CRISPR-Cas. 利用 CRISPR-Cas 生成猪繁殖与呼吸综合征病毒抗性猪的商业规模创始人种群。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-02-01 DOI: 10.1089/crispr.2023.0061

Brian T Burger, Benjamin P Beaton, Matthew A Campbell, Benjamin T Brett, Melissa S Rohrer, Sarah Plummer, Dylan Barnes, Ke Jiang, Sudhir Naswa, Jeremy Lange, Alina Ott, Elizabeth Alger, Gonzalo Rincon, Steven Rounsley, Jeff Betthauser, Namdori R Mtango, Joshua A Benne, Jessica Hammerand, Codie J Durfee, Marisa L Rotolo, Peter Cameron, Alexandra M Lied, Matthew J Irby, David B Nyer, Chris K Fuller, Scott Gradia, Steven B Kanner, Ki-Eun Park, Jerel Waters, Sean Simpson, Bhanu P Telugu, Brianna C Salgado, Alberto Brandariz-Nuñez, Raymond R R Rowland, Matt Culbertson, Elena Rice, A Mark Cigan

{"title":"Generation of a Commercial-Scale Founder Population of Porcine Reproductive and Respiratory Syndrome Virus Resistant Pigs Using CRISPR-Cas.","authors":"Brian T Burger, Benjamin P Beaton, Matthew A Campbell, Benjamin T Brett, Melissa S Rohrer, Sarah Plummer, Dylan Barnes, Ke Jiang, Sudhir Naswa, Jeremy Lange, Alina Ott, Elizabeth Alger, Gonzalo Rincon, Steven Rounsley, Jeff Betthauser, Namdori R Mtango, Joshua A Benne, Jessica Hammerand, Codie J Durfee, Marisa L Rotolo, Peter Cameron, Alexandra M Lied, Matthew J Irby, David B Nyer, Chris K Fuller, Scott Gradia, Steven B Kanner, Ki-Eun Park, Jerel Waters, Sean Simpson, Bhanu P Telugu, Brianna C Salgado, Alberto Brandariz-Nuñez, Raymond R R Rowland, Matt Culbertson, Elena Rice, A Mark Cigan","doi":"10.1089/crispr.2023.0061","DOIUrl":"10.1089/crispr.2023.0061","url":null,"abstract":"Disease resistance genes in livestock provide health benefits to animals and opportunities for farmers to meet the growing demand for affordable, high-quality protein. Previously, researchers used gene editing to modify the porcine CD163 gene and demonstrated resistance to a harmful virus that causes porcine reproductive and respiratory syndrome (PRRS). To maximize potential benefits, this disease resistance trait needs to be present in commercially relevant breeding populations for multiplication and distribution of pigs. Toward this goal, a first-of-its-kind, scaled gene editing program was established to introduce a single modified CD163 allele into four genetically diverse, elite porcine lines. This effort produced healthy pigs that resisted PRRS virus infection as determined by macrophage and animal challenges. This founder population will be used for additional disease and trait testing, multiplication, and commercial distribution upon regulatory approval. Applying CRISPR-Cas to eliminate a viral disease represents a major step toward improving animal health.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":"7 1","pages":"12-28"},"PeriodicalIF":3.7,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139731029","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

"CRISPR" Mutations: Inaccurate Linguistic Variations and Misrepresentation of the CRISPR Acronym. "CRISPR "突变：不准确的语言变化和对 CRISPR 首字母缩写词的错误表述。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-02-01 DOI: 10.1089/crispr.2024.0005

Jaime A Teixeira da

引用次数: 0

Generation of Human Isogenic Induced Pluripotent Stem Cell Lines with CRISPR Prime Editing. 利用 CRISPR 基因编辑技术生成人类同源诱导多能干细胞系。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2024-02-01 DOI: 10.1089/crispr.2023.0066

Lori L Bonnycastle, Amy J Swift, Erin C Mansell, Angela Lee, Elizabeth Winnicki, Elizabeth S Li, Catherine C Robertson, Victoria A Parsons, Trung Huynh, Chad Krilow, Karen L Mohlke, Michael R Erdos, Narisu Narisu, Francis S Collins

{"title":"Generation of Human Isogenic Induced Pluripotent Stem Cell Lines with CRISPR Prime Editing.","authors":"Lori L Bonnycastle, Amy J Swift, Erin C Mansell, Angela Lee, Elizabeth Winnicki, Elizabeth S Li, Catherine C Robertson, Victoria A Parsons, Trung Huynh, Chad Krilow, Karen L Mohlke, Michael R Erdos, Narisu Narisu, Francis S Collins","doi":"10.1089/crispr.2023.0066","DOIUrl":"10.1089/crispr.2023.0066","url":null,"abstract":"We developed an efficient CRISPR prime editing protocol and generated isogenic-induced pluripotent stem cell (iPSC) lines carrying heterozygous or homozygous alleles for putatively causal single nucleotide variants at six type 2 diabetes loci (ABCC8, MTNR1B, TCF7L2, HNF4A, CAMK1D, and GCK). Our two-step sequence-based approach to first identify transfected cell pools with the highest fraction of edited cells significantly reduced the downstream efforts to isolate single clones of edited cells. We found that prime editing can make targeted genetic changes in iPSC and optimization of system components and guide RNA designs that were critical to achieve acceptable efficiency. Systems utilizing PEmax, epegRNA modifications, and MLH1dn provided significant benefit, producing editing efficiencies of 36-73%. Editing success and pegRNA design optimization required for each variant differed depending on the sequence at the target site. With attention to design, prime editing is a promising approach to generate isogenic iPSC lines, enabling the study of specific genetic changes in a common genetic background.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":"7 1","pages":"53-67"},"PeriodicalIF":3.7,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10880268/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139731030","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