A5082977464- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
- Advanced biosensing and bioanalysis techniques
- Virus-based gene therapy research
- RNA regulation and disease
- Cytomegalovirus and herpesvirus research
- Mosquito-borne diseases and control
- Insect symbiosis and bacterial influences
- Genetics, Aging, and Longevity in Model Organisms
- RNA modifications and cancer
- Plant Virus Research Studies
- Chemical Synthesis and Analysis
- RNA Research and Splicing
- Vibrio bacteria research studies
- Retinal Development and Disorders
- Genetics and Neurodevelopmental Disorders
- Monoclonal and Polyclonal Antibodies Research
- Pluripotent Stem Cells Research
- Machine Learning in Bioinformatics
- Genomics and Rare Diseases
- Systemic Lupus Erythematosus Research
- Electrocatalysts for Energy Conversion
- Ubiquitin and proteasome pathways
- HIV Research and Treatment
- MicroRNA in disease regulation
University of Massachusetts Chan Medical School
2017-2023
Institut thématique Immunologie, inflammation, infectiologie et microbiologie
2017
Purdue University West Lafayette
2015
Lawrence University
2011
Abstract Prime editors (PEs) mediate genome modification without utilizing double-stranded DNA breaks or exogenous donor as a template. PEs facilitate nucleotide substitutions local insertions deletions within the based on template sequence encoded prime editing guide RNA (pegRNA). However, efficacy of in adult mice has not been established. Here we report an NLS-optimized SpCas9-based editor that improves efficiency both fluorescent reporter cells and at endogenous loci cultured cell lines....
The development of CRISPR genome editing has transformed biomedical research. Most applications reported thus far rely upon the Cas9 protein from Streptococcus pyogenes SF370 (SpyCas9). With many RNA guides, wildtype SpyCas9 can induce significant levels unintended mutations at near-cognate sites, necessitating substantial efforts toward strategies to minimize off-target activity. Although genome-editing potential thousands other orthologs remains largely untapped, it is not known how will...
Clustered, regularly interspaced, short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) have recently opened a new avenue for gene therapy. Cas9 nuclease guided by single-guide RNA (sgRNA) has been extensively used genome editing. Currently, three orthologs adapted in vivo engineering applications: Streptococcus pyogenes (SpyCas9), Staphylococcus aureus (SauCas9), Campylobacter jejuni (CjeCas9). However, additional editing platforms are needed, part to enable greater range...
Abstract RNA-based drugs depend on chemical modifications to increase potency and decrease immunogenicity in vivo. Chemical modification will likely improve the guide RNAs involved CRISPR-Cas9-based therapeutics as well. Cas9 orthologs are RNA-guided microbial effectors that cleave DNA. Here, we explore at all positions of crRNA tracrRNA cofactor. We identify several heavily modified versions more potent than their unmodified counterparts. In addition, describe fully chemically crRNAs...
As one of their countermeasures against CRISPR-Cas immunity, bacteriophages have evolved natural inhibitors known as anti-CRISPR (Acr) proteins. Despite the existence such examples for type II systems, we currently know relatively little about breadth Cas9 inhibitors, and most direct targets are uncharacterized. In this work identify two new II-C anti-CRISPRs cognate orthologs, validate functionality in vitro bacteria, define inhibitory spectrum a panel demonstrate that they act before DNA...
Adeno-associated virus (AAV) vectors are important delivery platforms for therapeutic genome editing but severely constrained by cargo limits. Simultaneous of multiple can limit dose and efficacy increase safety risks. Here, we describe single-vector, ~4.8-kb AAV that express Nme2Cas9 either two sgRNAs segmental deletions, or a single sgRNA with homology-directed repair (HDR) template. We also use anti-CRISPR proteins to enable production self-inactivate via cleavage. further introduce...
Nuclease-directed genome editing is a powerful tool for investigating physiology and has great promise as therapeutic approach to correct mutations that cause disease. In its most precise form, can use cellular homology-directed repair (HDR) pathways insert information from an exogenously supplied DNA-repair template (donor) directly into targeted genomic location. Unfortunately, particularly long insertions, toxicity delivery considerations associated with DNA limit HDR efficacy. Here, we...
The crystal structure of the hammerhead ribozyme bound to pentavalent transition state analogue vanadate reveals significant rearrangements relative previously determined structures. active site contracts, bringing G10.1 closer cleavage and repositioning a divalent metal ion such that it could, ultimately, interact directly with scissile phosphate. This could also position water molecule serve as general acid in reaction. A second is observed coordinated O6 G12. well-placed help tune pKA On...
The hammerhead ribozyme is a self-cleaving RNA broadly dispersed across all kingdoms of life. Although it was the first small, nucleolytic ribozymes discovered, mechanism by which catalyzes its reaction remains elusive. nucleobase G12 well positioned to be general base, but unclear if or how this guanine base becomes activated for proton transfer. Metal ions have been implicated in chemical mechanism, no interactions between divalent metal and cleavage site observed crystallographically. To...
While genome editing has been revolutionized by the advent of CRISPR-based nucleases, difficulties in achieving efficient, nuclease-mediated, homology-directed repair (HDR) still limit many applications. Commonly used DNA donors such as plasmids suffer from low HDR efficiencies cell types, well integration at unintended sites. In contrast, single-stranded (ssDNA) can produce efficient with minimal off-target integration. this study, we describe use ssDNA phage to efficiently and...
CRISPR-Cas9 systems provide powerful tools for genome editing. However, optimal employment of this technology will require control Cas9 activity so that the timing, tissue specificity, and accuracy editing may be precisely modulated. Anti-CRISPR proteins, which are small, naturally occurring inhibitors CRISPR-Cas systems, well suited purpose. A number anti-CRISPR proteins have been shown to potently inhibit subgroups but their maximal inhibitory is generally restricted specific homologs....
<h3>Background</h3> Precision medicine aims at providing intervention based on clinical and molecular stratification of patients, is an important approach for targeting heterogeneous diseases. A diverse autoimmune disease systemic lupus erythematosus (SLE), where dysregulation several immune processes affects multiple organs. Fundamental targeted treatment such a the identification biomarkers predictive biological basis phenotypes. <h3>Objectives</h3> Despite recent progress, few markers SLE...
Abstract Nuclease-directed genome editing is a powerful tool for investigating physiology and has great promise as therapeutic approach to correct mutations that cause disease. In its most precise form, can use cellular homology-directed repair (HDR) pathways insert information from an exogenously supplied DNA template (donor) directly into targeted genomic location. Unfortunately, particularly long insertions, toxicity delivery considerations associated with limit HDR efficacy. Here, we...
Abstract While genome editing has been revolutionized by the advent of CRISPR-based nucleases, difficulties in achieving efficient, nuclease-mediated, homology-directed repair (HDR) still limit many applications. Commonly used DNA donors such as plasmids suffer from low HDR efficiencies cell types, well integration at unintended sites. In contrast, single-stranded (ssDNA) can produce efficient with minimal off-target integration. Here, we describe use ssDNA phage to efficiently and...
ABSTRACT Background The development of CRISPR genome editing has transformed biomedical research. Most applications reported thus far rely upon the Cas9 protein from Streptococcus pyogenes SF370 (SpyCas9). With many RNA guides, wild-type SpyCas9 can induce significant levels unintended mutations at near-cognate sites, necessitating substantial efforts toward strategies to minimize off-target activity. Although genome-editing potential thousands other orthologs remains largely untapped, it is...
CRISPR-based genome-editing technologies, including nuclease editing, base and prime have recently revolutionized the development of therapeutics targeting disease-causing mutations. To advance assessment genome editing tools, a robust mouse model is valuable, particularly for evaluating
CRISPR-Cas systems are widely used for genome engineering technologies, and in their natural setting, they play crucial roles bacterial archaeal adaptive immunity, protecting against phages other mobile genetic elements. Previously we discovered bacteriophage-encoded Cas9-specific anti-CRISPR (Acr) proteins that serve as countermeasures host immunity by inactivating 1 . We hypothesized the evolutionary advantages conferred anti-CRISPRs would drive widespread occurrence of these nature 2–4...
CRISPR-Cas9 systems provide powerful tools for genome editing. However, optimal employment of this technology will require control Cas9 activity so that the timing, tissue specificity, and accuracy editing may be precisely modulated. Anti-CRISPR proteins, which are small naturally-occurring inhibitors CRISPR-Cas systems, well-suited purpose. A number anti-CRISPR proteins have been shown to potently inhibit subgroups but their maximal inhibitory is generally restricted specific homologs....
Abstract Prime editors (PEs) mediate genome modification without utilizing double-stranded DNA breaks or exogenous donor as a template. PEs facilitate nucleotide substitutions local insertions deletions within the based on template sequence encoded prime editing guide RNA (pegRNA). However, efficacy of in adult mice has not been established. Here we report an NLS-optimized SpCas9-based editor that improves efficiency both fluorescent reporter cells and at endogenous loci cultured cell lines....
RNA-based drugs depend on chemical modifications to increase potency and nuclease stability, decrease immunogenicity in vivo . Chemical modification will likely improve the guide RNAs involved CRISPR-Cas9-based therapeutics as well. Cas9 orthologs are RNA-guided microbial effectors that cleave DNA. No studies have yet explored at all positions of crRNA tracrRNA cofactor. Here, we identified several heavily-modified versions more potent than their unmodified counterparts. In addition,...
Abstract Adeno-associated virus (AAV) vectors are important delivery platforms for therapeutic genome editing but severely constrained by cargo limits, especially large effectors like Cas9s. Simultaneous of multiple can limit dose and efficacy increase safety risks. The use compact has enabled single-AAV Cas9s with 1-3 guides edits that end-joining repair pathways, many precise correct disease-causing mutations in vivo require homology-directed (HDR) templates. Here, we describe...