The Cas9 protein from the Streptococcus pyogenes CRISPR-Cas acquired immune system has been adapted for both RNA-guided genome editing and gene regulation in a variety of organisms, but it can mediate only a single activity at a time within any given cell. Here we characterize a set of fully orthogonal Cas9 proteins and demonstrate their ability to mediate simultaneous and independently targeted gene regulation and editing in bacteria and in human cells. We find that Cas9 orthologs display consistent patterns in their recognition of target sequences, and we identify an unexpectedly versatile Cas9 protein from Neisseria meningitidis. We provide a basal set of orthogonal RNA-guided proteins for controlling biological systems and establish a general methodology for characterizing additional proteins.
Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases
CRISPR-Cas systems in bacteria and archaea: versatile small RNAs for adaptive defense and regulation
Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria
Enhanced efficiency of human pluripotent stem cell genome editing through replacing TALENs with CRISPRs
One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering
Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system
CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering
Fusion of catalytically inactive Cas9 to FokI nuclease improves the specificity of genome modification
Nuclease Target Site Selection for Maximizing On-target Activity and Minimizing Off-target Effects in Genome Editing
Highly efficient heritable plant genome engineering using Cas9 orthologues from Streptococcus thermophilus and Staphylococcus aureus
Selection-free gene repair after adenoviral vector transduction of designer nucleases: rescue of dystrophin synthesis in DMD muscle cell populations
Expanding the CRISPR imaging toolset with Staphylococcus aureus Cas9 for simultaneous imaging of multiple genomic loci
Quantitative Analyses of Core Promoters Enable Precise Engineering of Regulated Gene Expression in Mammalian Cells
Suppression of hepatitis B virus DNA accumulation in chronically infected cells using a bacterial CRISPR/Cas RNA-guided DNA endonuclease
Trans-spliced Cas9 allows cleavage of HBB and CCR5 genes in human cells using compact expression cassettes
The three major types of CRISPR-Cas systems function independently in CRISPR RNA biogenesis in Streptococcus thermophilus
CRISPathBrick: Modular Combinatorial Assembly of Type II-A CRISPR Arrays for dCas9-Mediated Multiplex Transcriptional Repression in E. coli
Characterization of Staphylococcus aureus Cas9: a smaller Cas9 for all-in-one adeno-associated virus delivery and paired nickase applications
Precision metabolic engineering: The design of responsive, selective, and controllable metabolic systems
Conditionally Stabilized dCas9 Activator for Controlling Gene Expression in Human Cell Reprogramming and Differentiation
How the Knowledge of Interactions between Meningococcus and the Human Immune System Has Been Used to Prepare Effective Neisseria meningitidis Vaccines
Efficient inversions and duplications of mammalian regulatory DNA elements and gene clusters by CRISPR/Cas9
Controlling gene networks and cell fate with precision-targeted DNA-binding proteins and small-molecule-based genome readers
Multiplexed and programmable regulation of gene networks with an integrated RNA and CRISPR/Cas toolkit in human cells
Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers
Bacterial CRISPR/Cas DNA endonucleases: A revolutionary technology that could dramatically impact viral research and treatment
Multiplex CRISPR/Cas9-based genome editing for correction of dystrophin mutations that cause Duchenne muscular dystrophy
A programmable Cas9-serine recombinase fusion protein that operates on DNA sequences in mammalian cells
Cpf1 nucleases demonstrate robust activity to induce DNA modification by exploiting homology directed repair pathways in mammalian cells
CRISPRseek: a bioconductor package to identify target-specific guide RNAs for CRISPR-Cas9 genome-editing systems
A Biophysical Model of CRISPR/Cas9 Activity for Rational Design of Genome Editing and Gene Regulation
Programmable transcriptional repression in mycobacteria using an orthogonal CRISPR interference platform
Variant-aware saturating mutagenesis using multiple Cas9 nucleases identifies regulatory elements at trait-associated loci
Genome editing approaches: manipulating of lovastatin and taxol synthesis of filamentous fungi by CRISPR/Cas9 system
Efficient chromosomal gene modification with CRISPR/cas9 and PCR-based homologous recombination donors in cultured Drosophila cells
Hit-and-run epigenetic editing prevents senescence entry in primary breast cells from healthy donors
Inactivation of the human papillomavirus E6 or E7 gene in cervical carcinoma cells by using a bacterial CRISPR/Cas RNA-guided endonuclease
A revolutionary tool: CRISPR technology plays an important role in construction of intelligentized gene circuits
The CRISPR/Cas revolution continues: From efficient gene editing for crop breeding to plant synthetic biology
Orthologous CRISPR/Cas9 systems for specific and efficient degradation of covalently closed circular DNA of hepatitis B virus
Combining orthogonal CRISPR and CRISPRi systems for genome engineering and metabolic pathway modulation in Escherichia coli
Diversity, versatility and complexity of bacterial gene regulation mechanisms: opportunities and drawbacks for applications in synthetic biology
Highly efficient heritable targeted deletions of gene clusters and non-coding regulatory regions in Arabidopsis using CRISPR/Cas9
Next Generation Precision Medicine: CRISPR-mediated Genome Editing for the Treatment of Neurodegenerative Disorders
The rapidly advancing Class 2 CRISPR-Cas technologies: A customizable toolbox for molecular manipulations.
Genome-wide specificity of DNA binding, gene regulation, and chromatin remodeling by TALE- and CRISPR/Cas9-based transcriptional activators
Super-resolution microscopy reveals coupling between mammalian centriole subdistal appendages and distal appendages.
CRISPR Tools for Physiology and Cell State Changes: Potential of Transcriptional Engineering and Epigenome Editing.
* CRISPR-Based Epigenome Editing of Cytokine Receptors for the Promotion of Cell Survival and Tissue Deposition in Inflammatory Environments
Guiding Lights in Genome Editing for Inherited Retinal Disorders: Implications for Gene and Cell Therapy
Modular one-pot assembly of CRISPR arrays enables library generation and reveals factors influencing crRNA biogenesis
The Impact of CRISPR/Cas9 Technology on Cardiac Research: From Disease Modelling to Therapeutic Approaches
All-in-one adeno-associated virus delivery and genome editing by Neisseria meningitidis Cas9 in vivo
Mb- and FnCpf1 nucleases are active in mammalian cells: activities and PAM preferences of four wild-type Cpf1 nucleases and of their altered PAM specificity variants
Francisella novicida Cas9 interrogates genomic DNA with very high specificity and can be used for mammalian genome editing
Application and prospects of CRISPR/Cas9-based methods to trace defined genomic sequences in living and fixed plant cells.
The Development of an AAV-Based CRISPR SaCas9 Genome Editing System That Can Be Delivered to Neurons in vivo and Regulated via Doxycycline and Cre-Recombinase
New breeding technique "genome editing" for crop improvement: applications, potentials and challenges
CrisPam: SNP-Derived PAM Analysis Tool for Allele-Specific Targeting of Genetic Variants Using CRISPR-Cas Systems
SWITCH: a dynamic CRISPR tool for genome engineering and metabolic pathway control for cell factory construction in Saccharomyces cerevisiae
How Surrogate and Chemical Genetics in Model Organisms Can Suggest Therapies for Human Genetic Diseases
Cas Endonuclease Technology-A Quantum Leap in the Advancement of Barley and Wheat Genetic Engineering
A Cas12a ortholog with stringent PAM recognition followed by low off-target editing rates for genome editing.
CRISPR-Cas orthologues and variants: optimizing the repertoire, specificity and delivery of genome engineering tools
Biasing genome-editing events toward precise length deletions with an RNA-guided TevCas9 dual nuclease
Optogenetic control of Neisseria meningitidis Cas9 genome editing using an engineered, light-switchable anti-CRISPR protein.
Master Regulators and Cofactors of Human Neuronal Cell Fate Specification Identified by CRISPR Gene Activation Screens.
Covalent Modifications of the Bacteriophage Genome Confer a Degree of Resistance to Bacterial CRISPR Systems.
An extensive review to facilitate understanding of CRISPR technology as a gene editing possibility for enhanced therapeutic applications.
CRISPR/Cas: a Nobel Prize award-winning precise genome editing technology for gene therapy and crop improvement.
CRISPR Genome Editing & Therapy
CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on the application of this system for gene editing and therapy in human diseases.
Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.
CRISPR Ribonucleases Deactivation
CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.
Researcher Network:CZI Neurodegeneration Challenge
The Neurodegeneration Challenge Network aims to provide funding for and to bring together researchers studying neurodegenerative diseases. Find the latest research from the NDCN grantees here.