CRISPR Journal最新文献_第5页

Special Issue: CRISPR Trials. 特刊：CRISPR 试验。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2023-12-01 DOI: 10.1089/crispr.2023.29166.cfp2

Fyodor Urnov

引用次数: 0

The Expanding Dissemination and Distribution Patterns of Diverse CRISPR Plasmids by Addgene. 利用Addgene扩大CRISPR质粒的传播和分布模式。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2023-12-01 Epub Date: 2023-11-22 DOI: 10.1089/crispr.2023.0059

Brook Pyhtila, Seth Kasowitz, Rachel Leeson, Rodolphe Barrangou

{"title":"The Expanding Dissemination and Distribution Patterns of Diverse CRISPR Plasmids by Addgene.","authors":"Brook Pyhtila, Seth Kasowitz, Rachel Leeson, Rodolphe Barrangou","doi":"10.1089/crispr.2023.0059","DOIUrl":"10.1089/crispr.2023.0059","url":null,"abstract":"CRISPR-based technologies have rapidly enabled the democratization of genome editing in academic institutions through distribution by Addgene over the past decade. Recently, several distribution milestones have been reached, with a collection of >15,000 plasmids deposited by >1,000 laboratories spanning ∼40 countries now shipped 300,000 times to ∼5,000 organizations traversing ∼100 countries. Yet, both deposits of and requests for CRISPR plasmids continue to rise for this disruptive technology. Distribution patterns revealed robust demand for three distinct classes of CRISPR effectors, namely nucleases (e.g., Cas9 and Cas12), modulators (deactivated CRISPR nucleases fused to transcriptional regulators and epigenome modifiers), and chimeric effectors (Cas proteins fused to enzymes carrying out other activities such as deamination, reverse transcription, transposition, and integration). Yearly deposits over the past decade are requested in near-even proportions, reflecting continuous technological development and requests for novel constructs. Though it is unclear whether the slowing rate of requests is inherent to a pandemic operational lag or a transition from emerging to mature technology, it is noteworthy that the relative proportion of requests from plasmids deposited in the previous year remains stable, suggesting robust development of novel tools concurrent with continued adoption of editing, base editing, prime editing, and more. Predictably, most requested plasmids are designed for mammalian genome manipulation, presumably for medical research and human health pursuits, reflecting investments in therapeutic applications. Concurrently, requests for plant and microbial constructs are on the rise, especially in regions of the world more reliant on local agricultural inputs and focused on food and feed applications, illustrating continued diversification of genome editing applications.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10753985/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138447030","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

Engineered Antiviral Sensor Targets Infected Mosquitoes. 工程抗病毒传感器锁定受感染的蚊子

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2023-12-01 DOI: 10.1089/crispr.2023.0056

Elena Dalla Benetta, Adam J López-Denman, Hsing-Han Li, Reem A Masri, Daniel J Brogan, Michelle Bui, Ting Yang, Ming Li, Michael Dunn, Melissa J Klein, Sarah Jackson, Kyle Catalan, Kim R Blasdell, Priscilla Tng, Igor Antoshechkin, Luke S Alphey, Prasad N Paradkar, Omar S Akbari

{"title":"Engineered Antiviral Sensor Targets Infected Mosquitoes.","authors":"Elena Dalla Benetta, Adam J López-Denman, Hsing-Han Li, Reem A Masri, Daniel J Brogan, Michelle Bui, Ting Yang, Ming Li, Michael Dunn, Melissa J Klein, Sarah Jackson, Kyle Catalan, Kim R Blasdell, Priscilla Tng, Igor Antoshechkin, Luke S Alphey, Prasad N Paradkar, Omar S Akbari","doi":"10.1089/crispr.2023.0056","DOIUrl":"10.1089/crispr.2023.0056","url":null,"abstract":"Escalating vector disease burdens pose significant global health risks, as such innovative tools for targeting mosquitoes are critical. CRISPR-Cas technologies have played a crucial role in developing powerful tools for genome manipulation in various eukaryotic organisms. Although considerable efforts have focused on utilizing class II type II CRISPR-Cas9 systems for DNA targeting, these modalities are unable to target RNA molecules, limiting their utility against RNA viruses. Recently, the Cas13 family has emerged as an efficient tool for RNA targeting; however, the application of this technique in mosquitoes, particularly Aedes aegypti, has yet to be fully realized. In this study, we engineered an antiviral strategy termed REAPER (vRNA Expression Activates Poisonous Effector Ribonuclease) that leverages the programmable RNA-targeting capabilities of CRISPR-Cas13 and its potent collateral activity. REAPER remains concealed within the mosquito until an infectious blood meal is uptaken. Upon target viral RNA infection, REAPER activates, triggering programmed destruction of its target arbovirus such as chikungunya. Consequently, Cas13-mediated RNA targeting significantly reduces viral replication and viral prevalence of infection, and its promiscuous collateral activity can even kill infected mosquitoes within a few days. This innovative REAPER technology adds to an arsenal of effective molecular genetic tools to combat mosquito virus transmission.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11085028/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138807971","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

CRISPRi-Mediated Treatment of Dominant Rhodopsin-Associated Retinitis Pigmentosa. CRISPRi-Mediated Treatment of Dominant Rhodopsin-Associated Retinitis Pigmentosa.

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2023-12-01 DOI: 10.1089/crispr.2023.0039

Erin R Burnight, Luke A Wiley, Nathaniel K Mullin, Malavika K Adur, Mallory J Lang, Cathryn M Cranston, Chunhua Jiao, Stephen R Russell, Elliot H Sohn, Ian C Han, Jason W Ross, Edwin M Stone, Robert F Mullins, Budd A Tucker

{"title":"CRISPRi-Mediated Treatment of Dominant Rhodopsin-Associated Retinitis Pigmentosa.","authors":"Erin R Burnight, Luke A Wiley, Nathaniel K Mullin, Malavika K Adur, Mallory J Lang, Cathryn M Cranston, Chunhua Jiao, Stephen R Russell, Elliot H Sohn, Ian C Han, Jason W Ross, Edwin M Stone, Robert F Mullins, Budd A Tucker","doi":"10.1089/crispr.2023.0039","DOIUrl":"10.1089/crispr.2023.0039","url":null,"abstract":"Rhodopsin (RHO) mutations such as Pro23His are the leading cause of dominantly inherited retinitis pigmentosa in North America. As with other dominant retinal dystrophies, these mutations lead to production of a toxic protein product, and treatment will require knockdown of the mutant allele. The purpose of this study was to develop a CRISPR-Cas9-mediated transcriptional repression strategy using catalytically inactive Staphylococcus aureus Cas9 (dCas9) fused to the Krüppel-associated box (KRAB) transcriptional repressor domain. Using a reporter construct carrying green fluorescent protein (GFP) cloned downstream of the RHO promoter fragment (nucleotides -1403 to +73), we demonstrate a ∼74-84% reduction in RHO promoter activity in RHOpCRISPRi-treated versus plasmid-only controls. After subretinal transduction of human retinal explants and transgenic Pro23His mutant pigs, significant knockdown of rhodopsin protein was achieved. Suppression of mutant transgene in vivo was associated with a reduction in endoplasmic reticulum (ER) stress and apoptosis markers and preservation of photoreceptor cell layer thickness.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11304754/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138807791","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 Fluorescent Reporter Mouse for In Vivo Assessment of Genome Editing with Diverse Cas Nucleases and Prime Editors. 一种荧光报告小鼠，用于在体内评估使用多种 Cas 核酸酶和主编辑器进行基因组编辑的效果。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2023-12-01 DOI: 10.1089/crispr.2023.0048

Zexiang Chen, Suet-Yan Kwan, Aamir Mir, Max Hazeltine, Minwook Shin, Shun-Qing Liang, Io Long Chan, Karen Kelly, Krishna S Ghanta, Nicholas Gaston, Yueying Cao, Jun Xie, Guangping Gao, Wen Xue, Erik J Sontheimer, Jonathan K Watts

引用次数: 0

First Drug Approval Rises the CRISPR Tide. 首次药物批准掀起 CRISPR 浪潮。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2023-12-01 Epub Date: 2023-12-05 DOI: 10.1089/crispr.2023.29168.editorial

Rodolphe Barrangou

引用次数: 0

Correction to: APOBEC Reporter Systems for Evaluating diNucleotide Editing Levels by Rieffer et al. The CRISPR Journal, 2023;6(5):430-446; DOI: 10.1089/crispr.2023.0027. Rieffer等人对《评估双核核苷酸编辑水平的APOBEC报告系统》的更正。CRISPR期刊，2023；6（5）：430-446；DOI:10.1089/crispr.2023.0027。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2023-12-01 Epub Date: 2023-11-08 DOI: 10.1089/crispr.2023.0027.correx

引用次数: 0

Modified Cas9-Guided Oxford Nanopore Technology Sequencing Uncovers Single and Multiple Transgene Insertion Sites in a Zebrafish Melanoma Model. 改良 Cas9 引导的牛津纳米孔技术测序发现斑马鱼黑色素瘤模型中的单个和多个转基因插入位点。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2023-12-01 DOI: 10.1089/crispr.2023.0062

Raffaella De Paolo, Uday Munagala, Francesco Cucco, Samanta Sarti, Letizia Pitto, Filippo Martignano, Silvestro G Conticello, Laura Poliseno

引用次数: 0

A PCR-Induced Mutagenesis-Restriction Fragment Length Polymorphism Method for the Detection of CRISPR-Induced Indels. 用于检测 CRISPR 诱导的 Indels 的 PCR 诱导突变-限制性片段长度多态性方法。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2023-12-01 Epub Date: 2023-12-05 DOI: 10.1089/crispr.2023.0047

Lydia Angelopoulou, Electra Stylianopoulou, Konstantinos Tegopoulos, Ioanna Farmakioti, Maria Grigoriou, George Skavdis

引用次数: 0

Differential Divalent Metal Binding by SpyCas9's RuvC Active Site Contributes to Nonspecific DNA Cleavage. SpyCas9的RuvC活性位点与二价金属的不同结合导致了非特异性DNA裂解。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2023-12-01 DOI: 10.1089/crispr.2023.0022

Sydney N Newsom, Duen-Shian Wang, Saadi Rostami, Isabelle Schuster, Hari Priya Parameshwaran, Yadin G Joseph, Peter Z Qin, Jin Liu, Rakhi Rajan

{"title":"Differential Divalent Metal Binding by SpyCas9's RuvC Active Site Contributes to Nonspecific DNA Cleavage.","authors":"Sydney N Newsom, Duen-Shian Wang, Saadi Rostami, Isabelle Schuster, Hari Priya Parameshwaran, Yadin G Joseph, Peter Z Qin, Jin Liu, Rakhi Rajan","doi":"10.1089/crispr.2023.0022","DOIUrl":"10.1089/crispr.2023.0022","url":null,"abstract":"To protect against mobile genetic elements (MGEs), some bacteria and archaea have clustered regularly interspaced short palindromic repeats-CRISPR associated (CRISPR-Cas) adaptive immune systems. CRISPR RNAs (crRNAs) bound to Cas nucleases hybridize to MGEs based on sequence complementarity to guide the nucleases to cleave the MGEs. This programmable DNA cleavage has been harnessed for gene editing. Safety concerns include off-target and guide RNA (gRNA)-free DNA cleavages, both of which are observed in the Cas nuclease commonly used for gene editing, Streptococcus pyogenes Cas9 (SpyCas9). We developed a SpyCas9 variant (SpyCas9H982A) devoid of gRNA-free DNA cleavage activity that is more selective for on-target cleavage. The H982A substitution in the metal-dependent RuvC active site reduces Mn2+-dependent gRNA-free DNA cleavage by ∼167-fold. Mechanistic molecular dynamics analysis shows that Mn2+, but not Mg2+, produces a gRNA-free DNA cleavage competent state that is disrupted by the H982A substitution. Our study demonstrates the feasibility of modulating cation:protein interactions to engineer safer gene editing tools.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10753984/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138807848","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