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Expanding the Toolkit for Genome Editing in a Disease Vector, Aedes aegypti: Transgenic Lines Expressing Cas9 and Single Guide RNA Induce Efficient Mutagenesis CRISPR J. (IF 5.343) Pub Date : 2021-01-15 Guan-Heng Zhu; Najla M. Albishi; Xien Chen; Rachel L. Brown; Subba Reddy Palli
CRISPR-Cas9 mediated genome editing methods are being used for the analysis of gene function. However, it is hard to identify gene knockout mutants for genes whose knockout does not cause distinct phenotypes. To overcome this issue in the disease vector, Aedes aegypti, a transgenic Cas9/single guide RNA (sgRNA) method, was used to knock out the eye marker gene, kynurenine 3-monooxygenase (kmo), and
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Heritable Human Genome Editing: The Public Engagement Imperative CRISPR J. (IF 5.343) Pub Date : 2020-12-17 Eli Y. Adashi; Michael M. Burgess; Simon Burall; I. Glenn Cohen; Leonard M. Fleck; John Harris; Soren Holm; Cristina Lafont; Jonathan D. Moreno; Michael A. Neblo; Simon J. Niemeyer; Eugene J. Rowe; Dietram A. Scheufele; Paul F. Tetsa; Effy Vayena; Richard P. Watermeyer; Archon Fung
In the view of many, heritable human genome editing (HHGE) harbors the remedial potential of ridding the world of deadly genetic diseases. A Hippocratic obligation, if there ever was one, HHGE is widely viewed as a life-sustaining proposition. The national go/no-go decision regarding the implementation of HHGE, however, must not, in the collective view of the authors, proceed absent thorough public
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Evaluation of Homology-Independent CRISPR-Cas9 Off-Target Assessment Methods CRISPR J. (IF 5.343) Pub Date : 2020-12-18 Hemangi G. Chaudhari; Jon Penterman; Holly J. Whitton; Sarah J. Spencer; Nicole Flanagan; Maria C. Lei Zhang; Elaine Huang; Aditya S. Khedkar; J. Mike Toomey; Courtney A. Shearer; Alexander W. Needham; Tony W. Ho; John D. Kulman; T.J. Cradick; Andrew Kernytsky
The ability to alter genomes specifically by CRISPR-Cas gene editing has revolutionized biological research, biotechnology, and medicine. Broad therapeutic application of this technology, however, will require thorough preclinical assessment of off-target editing by homology-based prediction coupled with reliable methods for detecting off-target editing. Several off-target site nomination assays exist
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Novel Type V-A CRISPR Effectors Are Active Nucleases with Expanded Targeting Capabilities CRISPR J. (IF 5.343) Pub Date : 2020-12-17 Daniela S. Aliaga Goltsman; Lisa M. Alexander; Audra E. Devoto; Justine B. Albers; Jason Liu; Cristina N. Butterfield; Christopher T. Brown; Brian C. Thomas
Cas12a enzymes are quickly being adopted for use in a variety of genome-editing applications. These programmable nucleases are part of adaptive microbial immune systems, the natural diversity of which has been largely unexplored. Here, we identified novel families of Type V-A CRISPR nucleases through a large-scale analysis of metagenomes collected from a variety of complex environments, and developed
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CRISPRCasTyper: Automated Identification, Annotation, and Classification of CRISPR-Cas Loci CRISPR J. (IF 5.343) Pub Date : 2020-12-18 Jakob Russel; Rafael Pinilla-Redondo; David Mayo-Muñoz; Shiraz A. Shah; Søren J. Sørensen
Automated classification of CRISPR-Cas systems has been challenged by their dynamic nature and expanding classification. Here, we developed CRISPRCasTyper, an automated tool with improved capabilities for identifying and typing CRISPR arrays and cas loci based on the latest nomenclature (44 subtypes/variants). As a novel feature, CRISPRCasTyper uses a machine learning approach to subtype CRISPR arrays
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A CRISPR-Cas9, Cre-lox, and Flp-FRT Cascade Strategy for the Precise and Efficient Integration of Exogenous DNA into Cellular Genomes CRISPR J. (IF 5.343) Pub Date : 2020-12-17 Jixue Li; Yanjie Li; Kevin M. Pawlik; Jill S. Napierala; Marek Napierala
We describe a protocol for the precise integration of exogenous DNA into user-defined genomic loci in cultured cells. This strategy first introduces a promoter and a lox site to a specific location via a Cas9-induced double-strand break. Second, a gene of interest (GOI) is inserted into the lox site via Cre-lox recombination. Upon correct insertion, a cis-linked antibiotic resistance gene will be expressed
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CRISPR-Assisted DNA Detection: A Novel dCas9-Based DNA Detection Technique CRISPR J. (IF 5.343) Pub Date : 2020-12-17 Xinhui Xu; Tao Luo; Jinliang Gao; Na Lin; Weiwei Li; Xinyi Xia; Jinke Wang
Nucleic acid detection techniques are always critical to diagnosis, especially in the background of the present coronavirus disease 2019 pandemic. Simple and rapid detection techniques with high sensitivity and specificity are always urgently needed. However, current nucleic acid detection techniques are still limited by traditional amplification and hybridization. To overcome this limitation, here
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Improving FnCas12a Genome Editing by Exonuclease Fusion CRISPR J. (IF 5.343) Pub Date : 2020-12-18 Yongqiang Wu; Qichen Yuan; Yufeng Zhu; Xiang Gao; Jiabao Song; Ziru Yin
Among current reported Cas12a orthologs, Francisella novicida Cas12a (FnCas12a) is less restricted by protospacer adjacent motif (PAM). However, the activity of FnCas12a nuclease is relatively low or undetectable in human cells, limiting its application as desirable genome engineering tools. Here, we describe TEXT (Tethering EXonuclease T5 with FnCas12a)—a fusion strategy that significantly increased
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AsCRISPR: A Web Server for Allele-Specific Single Guide RNA Design in Precision Medicine CRISPR J. (IF 5.343) Pub Date : 2020-12-18 Guihu Zhao; Jinchen Li; Yu Tang
Allele-specific genomic targeting by CRISPR is a versatile strategy that has been increasingly exploited not only in treating inherited dominant diseases and mutation-driven cancers, but also in other important fields such as genome imprinting, haploinsufficiency, and genome loci imaging. Despite its tremendous utilities, few bioinformatic tools have been implemented for the allele-specific purpose
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One-Step Homology Mediated CRISPR-Cas Editing in Zygotes for Generating Genome Edited Cattle CRISPR J. (IF 5.343) Pub Date : 2020-12-17 Ki-Eun Park; Juli Foster Frey; Jerel Waters; Sean G. Simpson; Chris Coutu; Sarah Plummer; Matthew Campbell; David M. Donovan; Bhanu P. Telugu
Selective breeding and genetic modification have been the cornerstone of animal agriculture. However, the current strategy of breeding animals over multiple generations to introgress novel alleles is not practical in addressing global challenges such as climate change, pandemics, and the predicted need to feed a population of 9 billion by 2050. Consequently, genome editing in zygotes to allow for seamless
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CRISPR Arrays Away from cas Genes CRISPR J. (IF 5.343) Pub Date : 2020-12-18 Sergey A. Shmakov; Irina Utkina; Yuri I. Wolf; Kira S. Makarova; Konstantin V. Severinov; Eugene V. Koonin
CRISPR-Cas systems typically consist of a CRISPR array and cas genes that are organized in one or more operons. However, a substantial fraction of CRISPR arrays are not adjacent to cas genes. Definitive identification of such isolated CRISPR arrays runs into the problem of false-positives, with unrelated types of repetitive sequences mimicking CRISPR. We developed a computational pipeline to eliminate
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CRISPR-Cas “Non-Target” Sites Inhibit On-Target Cutting Rates CRISPR J. (IF 5.343) Pub Date : 2020-12-18 Eirik A. Moreb; Mitchell Hutmacher; Michael D. Lynch
CRISPR-Cas systems have become ubiquitous for genome editing in eukaryotic as well as bacterial systems. Cas9 forms a complex with a guide RNA (gRNA) and searches DNA for a matching sequence (target site) next to a protospacer adjacent motif (PAM). Once found, Cas9 cuts the DNA. Cas9 is revolutionary for the ability to change the RNA sequence and target a new site easily. However, while algorithms
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Regenerating Urethral Striated Muscle by CRISPRi/dCas9-KRAB-Mediated Myostatin Silencing for Obesity-Associated Stress Urinary Incontinence CRISPR J. (IF 5.343) Pub Date : 2020-12-18 Huixing Yuan; Yajun Ruan; Yan Tan; Amanda B. Reed-Maldonado; Yinwei Chen; Dehua Zhao; Zhao Wang; Feng Zhou; Dongyi Peng; Lia Banie; Guifang Wang; Jihong Liu; Guiting Lin; Lei S. Qi; Tom F. Lue
Overweight females are prone to obesity-associated stress urinary incontinence (OA-SUI), and there are no definitive medical therapies for this common urologic condition. This study was designed to test the hypothesis that regenerative therapy to restore urethral striated muscle (stM) and pelvic floor muscles might represent a valuable therapeutic approach. For the in vitro experiment, single-guide
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Reactions to the National Academies/Royal Society Report on Heritable Human Genome Editing CRISPR J. (IF 5.343) Pub Date : 2020-10-20 Misha Angrist; Rodolphe Barrangou; Françoise Baylis; Carolyn Brokowski; Gaetan Burgio; Arthur Caplan; Carolyn Riley Chapman; George M. Church; Robert Cook-Deegan; Bryan Cwik; Jennifer A. Doudna; John H. Evans; Henry T. Greely; Laura Hercher; J. Benjamin Hurlbut; Richard O. Hynes; Tetsuya Ishii; Samira Kiani; LaTasha Hoskins Lee; Guillaume Levrier; David R. Liu; Jeantine E. Lunshof; Kerry Lynn Macintosh;
In September 2020, a detailed report on Heritable Human Genome Editing was published. The report offers a translational pathway for the limited approval of germline editing under limited circumstances and assuming various criteria have been met. In this perspective, some three dozen experts from the fields of genome editing, medicine, bioethics, law, and related fields offer their candid reactions
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CRISPR-Cas Activators for Engineering Gene Expression in Higher Eukaryotes CRISPR J. (IF 5.343) Pub Date : 2020-10-20 J. Armando Casas-Mollano; Matthew H. Zinselmeier; Samuel E. Erickson; Michael J. Smanski
CRISPR-Cas-based transcriptional activators allow genetic engineers to specifically induce expression of one or many target genes in trans. Here we review the many design variations of these versatile tools and compare their effectiveness in different eukaryotic systems. Lastly, we highlight several applications of programmable transcriptional activation to interrogate and engineer complex biological
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Human Germline and Heritable Genome Editing: The Global Policy Landscape CRISPR J. (IF 5.343) Pub Date : 2020-10-20 Françoise Baylis; Marcy Darnovsky; Katie Hasson; Timothy M. Krahn
Discussions and debates about the governance of human germline and heritable genome editing should be informed by a clear and accurate understanding of the global policy landscape. This policy survey of 106 countries yields significant new data. A large majority of countries (96 out of 106) surveyed have policy documents—legislation, regulations, guidelines, codes, and international treaties—relevant
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Reproducible Antigen Recognition by the Type I-F CRISPR-Cas System CRISPR J. (IF 5.343) Pub Date : 2020-10-20 Tanner Wiegand; Ekaterina Semenova; Anna Shiriaeva; Ivan Fedorov; Kirill Datsenko; Konstantin Severinov; Blake Wiedenheft
CRISPR-associated proteins 1 and 2 (Cas1–2) are necessary and sufficient for new spacer acquisition in some CRISPR-Cas systems (e.g., type I-E), but adaptation in other systems (e.g., type II-A) involves the crRNA-guided surveillance complex. Here we show that the type I-F Cas1–2/3 proteins are necessary and sufficient to produce low levels of spacer acquisition, but the presence of the type I-F crRNA-guided
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Progress Toward Zygotic and Germline Gene Drives in Mice CRISPR J. (IF 5.343) Pub Date : 2020-10-20 Chandran Pfitzner; Melissa A. White; Sandra G. Piltz; Michaela Scherer; Fatwa Adikusuma; James N. Hughes; Paul Q. Thomas
CRISPR-based synthetic gene drives have the potential to deliver a more effective and humane method of invasive vertebrate pest control than current strategies. Relatively efficient CRISPR gene drive systems have been developed in insects and yeast but not in mammals. Here, we investigated the efficiency of CRISPR-Cas9-based gene drives in Mus musculus by constructing “split drive” systems where gRNA
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Engineered RNA-Interacting CRISPR Guide RNAs for Genetic Sensing and Diagnostics CRISPR J. (IF 5.343) Pub Date : 2020-10-20 Roberto Galizi; John N. Duncan; William Rostain; Charlotte M Quinn; Marko Storch; Manish Kushwaha; Alfonso Jaramillo
CRISPR guide RNAs (gRNAs) can be programmed with relative ease to allow the genetic editing of nearly any DNA or RNA sequence. Here, we propose novel molecular architectures to achieve RNA-dependent modulation of CRISPR activity in response to specific RNA molecules. We designed and tested, in both living Escherichia coli cells and cell-free assays for rapid prototyping, cis-repressed RNA-interacting
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In Vitro Validation of Transgene Expression in Gene-Edited Pigs Using CRISPR Transcriptional Activators CRISPR J. (IF 5.343) Pub Date : 2020-10-20 Kathryn M. Polkoff; Jaewook Chung; Sean G. Simpson; Katherine Gleason; Jorge A. Piedrahita
The use of CRISPR-Cas and RNA-guided endonucleases has drastically changed research strategies for understanding and exploiting gene function, particularly for the generation of gene-edited animal models. This has resulted in an explosion in the number of gene-edited species, including highly biomedically relevant pig models. However, even with error-free DNA insertion or deletion, edited genes are
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An Examination of Public Discourse on Human Gene Editing Using Natural Language Processing. CRISPR J. (IF 5.343) Pub Date : 2020-08-24 Micah Musser
This research aims to explore the different ways in which scientists, ethicists, journalists, and commissions speak to the public about new gene-editing technologies. The research collected more than 100,000 sentences from books, news articles, and reports written by these four types of authors, examined the relative distinctiveness of their speech, and compared the prevalence of key terms among the
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Sharing the CRISPR Toolbox with an Expanding Community. CRISPR J. (IF 5.343) Pub Date : 2020-08-24 Caroline M LaManna,Brook Pyhtila,Rodolphe Barrangou
Over the past 8 years, the widespread adoption of CRISPR-based technologies has fueled the global genome editing revolution. This platform is based on Cas molecular machines such as Cas9, Cas12, Cas13, as well as other CRISPR effector proteins that are able to alter the genome, transcriptome, and epigenome of virtually any species. Technological improvements have rendered these tools more efficient
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Translating CRISPR-Cas Therapeutics: Approaches and Challenges. CRISPR J. (IF 5.343) Pub Date : 2020-08-24 Lavina Sierra Tay,Nathan Palmer,Rebecca Panwala,Wei Leong Chew,Prashant Mali
CRISPR-Cas clinical trials have begun, offering a first glimpse at how DNA and RNA targeting could enable therapies for many genetic and epigenetic human diseases. The speedy progress of CRISPR-Cas from discovery and adoption to clinical use is built on decades of traditional gene therapy research and belies the multiple challenges that could derail the successful translation of these new modalities
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Promoter Orientation within an AAV-CRISPR Vector Affects Cas9 Expression and Gene Editing Efficiency. CRISPR J. (IF 5.343) Pub Date : 2020-08-24 Lewis E Fry,Caroline F Peddle,Marta Stevanovic,Alun R Barnard,Michelle E McClements,Robert E MacLaren
Adeno-associated virus (AAV) vectors have been widely adopted for delivery of CRISPR-Cas components, especially for therapeutic gene editing. For a single vector system, both the Cas9 and guide RNA (gRNA) are encoded within a single transgene, usually from separate promoters. Careful design of this bi-cistronic construct is required due to the minimal packaging capacity of AAV. We investigated how
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Low-Density Lipoprotein Receptor-Related Protein 5-Deficient Rats Have Reduced Bone Mass and Abnormal Development of the Retinal Vasculature. CRISPR J. (IF 5.343) Pub Date : 2020-08-24 John L Ubels,Cassandra R Diegel,Gabrielle E Foxa,Nicole J Ethen,Jonathan N Lensing,Zachary B Madaj,Bart O Williams
Humans carrying homozygous loss-of-function mutations in the Wnt co-receptor, low-density lipoprotein receptor–related protein 5 (LRP5), develop osteoporosis and a defective retinal vasculature known as familial exudative vitreoretinopathy (FEVR) due to disruption of the Wnt signaling pathway. The purpose of this study was to use CRISPR-Cas9–mediated gene editing to create strains of Lrp5-deficient
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Targeted RNA Knockdown by a Type III CRISPR-Cas Complex in Zebrafish. CRISPR J. (IF 5.343) Pub Date : 2020-08-24 Thomas Fricke,Dalia Smalakyte,Maciej Lapinski,Abhishek Pateria,Charles Weige,Michal Pastor,Agnieszka Kolano,Cecilia Winata,Virginijus Siksnys,Gintautas Tamulaitis,Matthias Bochtler
RNA interference is a powerful experimental tool for RNA knockdown, but not all organisms are amenable. Here, we provide a proof of principle demonstration that a type III Csm effector complex can be used for programmable mRNA transcript degradation in eukaryotes. In zebrafish, Streptococcus thermophilus Csm complex (StCsm) proved effective for knockdown of maternally expressed EGFP in germ cells of
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CRISPR Meets Proteomics: How IsoPlexis' Single Cell Data is Revolutionizing Therapeutic Genome Editing. CRISPR J. (IF 5.343) Pub Date : 2020-08-01
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A Navigation System for Base Editing: Are We There Yet? CRISPR J. (IF 5.343) Pub Date : 2020-08-01 Denis C Bauer,Laurence O W Wilson
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CRISPR-Free Mitochondrial DNA Base Editing. CRISPR J. (IF 5.343) Pub Date : 2020-08-01 Douglas C Wallace
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In Times Like These, We All Need a Moment of Science. CRISPR J. (IF 5.343) Pub Date : 2020-08-01 Rodolphe Barrangou
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Retrons: Complementing CRISPR in Phage Defense. CRISPR J. (IF 5.343) Pub Date : 2020-08-01 Karen L Maxwell
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What Do We (Not) Know About Global Views of Human Gene Editing? Insights and Blind Spots in the CRISPR Era CRISPR J. (IF 5.343) Pub Date : 2020-06-17 Emily L. Howell; Shiyu Yang; Becca Beets; Dominique Brossard; Dietram A. Scheufele; Michael A. Xenos
As research on human applications of CRISPR advances, researchers, advisory bodies, and other stakeholder organizations continue calling for global public discourses and engagement to shape the development of human gene editing (HGE). Research that captures public views and tests ways for engaging across viewpoints is vital for facilitating these discourses. Unfortunately, such research lags behind
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Unprecedented Diversity of Unique CRISPR-Cas-Related Systems and Cas1 Homologs in Asgard Archaea CRISPR J. (IF 5.343) Pub Date : 2020-06-17 Kira S. Makarova; Yuri I. Wolf; Sergey A. Shmakov; Yang Liu; Meng Li; Eugene V. Koonin
The principal function of archaeal and bacterial CRISPR-Cas systems is antivirus adaptive immunity. However, recent genome analyses identified a variety of derived CRISPR-Cas variants at least some of which appear to perform different functions. Here, we describe a unique repertoire of CRISPR-Cas-related systems that we discovered by searching archaeal metagenome-assemble genomes of the Asgard superphylum
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Programmable RNA Targeting Using CasRx in Flies. CRISPR J. (IF 5.343) Pub Date : 2020-06-17 Anna B Buchman,Dan J Brogan,Ruichen Sun,Ting Yang,Patrick D Hsu,Omar S Akbari
CRISPR-Cas genome editing technologies have revolutionized the fields of functional genetics and genome engineering, but with the recent discovery and optimization of RNA-targeting Cas ribonucleases, we may soon see a similar revolution in the study of RNA function and transcriptome engineering. However, to date, successful proof of principle for Cas ribonuclease RNA targeting in eukaryotic systems
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Detection and Modulation of DNA Translocations During Multi-Gene Genome Editing in T Cells. CRISPR J. (IF 5.343) Pub Date : 2020-06-17 Anne Bothmer,Kenneth W Gareau,Hayat S Abdulkerim,Frank Buquicchio,Lucas Cohen,Ramya Viswanathan,John A Zuris,Eugenio Marco,Cecilia A Fernandez,Vic E Myer,Cecilia Cotta-Ramusino
Multiplexed genome editing with DNA endonucleases has broad application, including for cellular therapies, but chromosomal translocations, natural byproducts of inducing simultaneous genomic breaks, have not been explored in detail. Here we apply various CRISPR-Cas nucleases to edit the T cell receptor alpha and beta 2 microglobulin genes in human primary T cells and comprehensively evaluate the frequency
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A Cellular Stress Response Induced by the CRISPR-dCas9 Activation System Is Not Heritable Through Cell Divisions CRISPR J. (IF 5.343) Pub Date : 2020-06-17 Andrew D. Johnston; Alali Abdulrazak; Hanae Sato; Shahina B. Maqbool; Masako Suzuki; John M. Greally; Claudia A. Simões-Pires
The CRISPR-Cas9 system can be modified to perform “epigenetic editing” by utilizing the catalytically inactive (dead) Cas9 (dCas9) to recruit regulatory proteins to specific genomic locations. In prior studies, epigenetic editing with multimers of the transactivator VP16 and guide RNAs (gRNAs) was found to cause adverse cellular responses. These side effects may confound studies inducing new cellular
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CRISPR-Cas9-Mediated Intersectional Knockout of Glycogen Synthase Kinase 3β in D2 Receptor-Expressing Medial Prefrontal Cortex Neurons Reveals Contributions to Emotional Regulation. CRISPR J. (IF 5.343) Pub Date : 2020-06-17 Jivan Khlghatyan,Jean-Martin Beaulieu
Glycogen synthase kinase 3β (GSK3β) activity is regulated by dopamine D2 receptor signaling and can be inhibited by psychoactive drugs in a D2 receptor–dependent manner. However, GSK3β is ubiquitously expressed in the brain, and D2 receptor–expressing cells are distributed as a mosaic in multiple cortical regions. This complicates the interrogation of GSK3β functions in cortical D2 cells in a circuit-defined
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A Flexible, Pooled CRISPR Library for Drug Development Screens CRISPR J. (IF 5.343) Pub Date : 2020-06-17 Maximilian Blanck; Milka B. Budnik-Zawilska; Steven R. Lenger; John E. McGonigle; Glynn R.A. Martin; Carlos le Sage; Steffen Lawo; Helen N. Pemberton; Gaganpreet S. Tiwana; David A. Sorrell; Benedict C.S. Cross
Functional genomic screening with CRISPR has provided a powerful and precise new way to interrogate the phenotypic consequences of gene manipulation in high-throughput, unbiased analyses. However, some experimental paradigms prove especially challenging and require carefully and appropriately adapted screening approaches. In particular, negative selection (or sensitivity) screening, often the most
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New Additions to the CRISPR Toolbox: CRISPR-CLONInG and CRISPR-CLIP for Donor Construction in Genome Editing. CRISPR J. (IF 5.343) Pub Date : 2020-04-21 Dorjee T N Shola,Chingwen Yang,Vhy-Shelta Kewaldar,Pradip Kar,Victor Bustos
CRISPR-Cas has proven to be the most versatile genetic tinkering system of our time, predominantly as a precision genome editing tool. Here, we demonstrate two additions to the repertoire of CRISPR's application for constructing donor DNA templates: CRISPR-CLONInG and CRISPR-CLIP. CRISPR-CLONInG (CRISPR-Cutting and Ligation Of Nucleic acid In vitro via Gibson) was devised to enable efficient cut-and-paste
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ErCas12a CRISPR-MAD7 for Model Generation in Human Cells, Mice, and Rats. CRISPR J. (IF 5.343) Pub Date : 2020-04-21 Zhenyi Liu,John A Schiel,Elena Maksimova,Žaklina Strezoska,Guojun Zhao,Emily M Anderson,Yumei Wu,Joe Warren,Angela Bartels,Anja van Brabant Smith,Chris E Lowe,Kevin P Forbes
MAD7 is an engineered class 2 type V-A CRISPR-Cas (Cas12a/Cpf1) system isolated from Eubacterium rectale. Analogous to Cas9, it is an RNA-guided nuclease with demonstrated gene editing activity in Escherichia coli and yeast cells. Here, we report that MAD7 is capable of generating indels and fluorescent gene tagging of endogenous genes in human HCT116 and U2OS cancer cell lines, respectively. In addition
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A Tandem Guide RNA-Based Strategy for Efficient CRISPR Gene Editing of Cell Populations with Low Heterogeneity of Edited Alleles. CRISPR J. (IF 5.343) Pub Date : 2020-04-21 Gérard Joberty,Maria Fälth-Savitski,Marcel Paulmann,Markus Bösche,Carola Doce,Aaron T Cheng,Gerard Drewes,Paola Grandi
CRISPR/Cas9–based gene knockouts (KOs) enable precise perturbation of target gene function in human cells, which is ideally assessed in an unbiased fashion by molecular omics readouts. Typically, this requires the lengthy process of isolating KO subclones. We show here that KO subclones are phenotypically heterogenous, regardless of the guide RNA used. We present an experimental strategy that avoids
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CRISPR-Cas9-Based Genome Editing in the Silverleaf Whitefly (Bemisia tabaci). CRISPR J. (IF 5.343) Pub Date : 2020-04-21 Chan C Heu,Francine M McCullough,Junbo Luan,Jason L Rasgon
Bemisia tabaci cryptic species Middle East-Asia Minor I (MEAM1) is a serious agricultural polyphagous insect pest and vector of numerous plant viruses, causing major worldwide economic losses. B. tabaci control is limited by lack of robust gene editing tools. Gene editing is difficult in B. tabaci due to small embryos that are technically challenging to inject and which have high mortality post injection
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Ethics and Global Governance of Human Germline Genome Editing: The Problem of Techno-Scientific Colonialist Paternalism. CRISPR J. (IF 5.343) Pub Date : 2020-04-21 Gabriela Arguedas-Ramírez
I want to enrich the debate about the ethics and governance of human germline editing (HGE) by emphasizing an underappreciated, yet important, set of concerns regarding exclusionary practices, norms, and efforts that impede a broader discussion about the subject. The possibility for establishing a binding, global, regulatory framework is influenced by economic and geopolitical factors as well as historical
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Updating the CRISPR Catalogue. CRISPR J. (IF 5.343) Pub Date : 2020-04-01 Yukti Dhingra,Dipali G Sashital
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COVID-19 and the CRISPR Community Response. CRISPR J. (IF 5.343) Pub Date : 2020-04-01 Kevin Davies,Rodolphe Barrangou
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The CRISPR-RNA World: An Interview with Martin Jínek. CRISPR J. (IF 5.343) Pub Date : 2020-04-01 Kevin Davies,Martin Jínek
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DNA Editing for Amyotrophic Lateral Sclerosis: Leading Off First Base. CRISPR J. (IF 5.343) Pub Date : 2020-04-01 Thomas J Cunningham,Elizabeth Fisher,Pietro Fratta,Jonathan D Gilthorpe
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CRISPR and the Law: A South African Perspective. CRISPR J. (IF 5.343) Pub Date : 2020-04-01 Sheetal Soni
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A CRISPR Odyssey into Cancer Immunotherapy. CRISPR J. (IF 5.343) Pub Date : 2020-04-01 Elizabeth Delgado,Samira Kiani
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Cautious Progress Toward Clinical Application of Human Gene Editing. CRISPR J. (IF 5.343) Pub Date : 2020-02-01 Sheetal Soni
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Ethical Considerations in Therapeutic Clinical Trials Involving Novel Human Germline-Editing Technology. CRISPR J. (IF 5.343) Pub Date : 2020-02-01 Carolyn Brokowski,Mazhar Adli
Much of the international community opposes editing the human germline. Yet, given enough experience to become better acquainted with strengths and limitations, prominent international figures are cautiously optimistic about using CRISPR-like novel technologies for clinical applications. Not only might such applications be morally (ethically) permissible, but clinical trials for therapeutic aims could
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Ordo-Responsibility for Germline Gene Editing. CRISPR J. (IF 5.343) Pub Date : 2020-02-01 Robert Ranisch,Tina Rudolph,Hans-Joachim Cremer,Nikolaus Knoepffler
The case of twins born with genes modified by He Jiankui highlights the need for international governance of germline gene editing (GGE). This article proposes a global framework that utilizes "ordo-responsibilities." This is a pragmatic ethical approach open to pluralism and grounded in principles of human dignity and human rights. Ordo-responsibility is pragmatic in (1) accepting generally available
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CRISPR-Cas9 Application in Canadian Public and Private Plant Breeding. CRISPR J. (IF 5.343) Pub Date : 2020-02-01 Savannah Gleim,Simona Lubieniechi,Stuart J Smyth
Plant-breeding technologies have expanded, accelerating breeding research beyond the confines of current regulations. The application of genome editing, such as CRISPR-Cas9, do not neatly fit into existing regulatory frameworks, creating uncertainty as to whether they can be regarded as conventionally developed varieties without further regulation. This research presents the current views of Canadian
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A Jumbo Formation in the Viral Game Plan. CRISPR J. (IF 5.343) Pub Date : 2020-02-01 Jeffrey K Cornuault,Sylvain Moineau
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Major Insights into Microbiology: An Interview with Luciano Marraffini. CRISPR J. (IF 5.343) Pub Date : 2020-02-01 Kevin Davies,Luciano Marraffini
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