![]() Genome editing of HBG1 and HBG2 to induce fetal hemoglobin. Editing aberrant splice sites efficiently restores β-globin expression in β-thalassemia. Selection-free genome editing of the sickle mutation in human adult hematopoietic stem/progenitor cells. CRISPR-Cas9 in vivo gene editing for transthyretin amyloidosis. CRISPR-engineered T cells in patients with refractory cancer. CRISPR-edited stem cells in a patient with HIV and acute lymphocytic leukemia. CRISPR/Cas9-mediated CCR5 ablation in human hematopoietic stem/progenitor cells confers HIV-1 resistance in vivo. A high-fidelity Cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human hematopoietic stem and progenitor cells. ![]() Highly parallel profiling of Cas9 variant specificity. Durable and robust fetal globin induction without Anemia in rhesus monkeys following autologous hematopoietic stem cell transplant with BCL11A erythroid enhancer editing. A machine learning approach for predicting CRISPR-Cas9 cleavage efficiencies and patterns underlying its mechanism of action. Expanded encyclopaedias of DNA elements in the human and mouse genomes. GENCODE reference annotation for the human and mouse genomes. CALITAS: A CRISPR-Cas-aware ALigner for In silico off-TArget Search. Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Highly efficient therapeutic gene editing of human hematopoietic stem cells. BCL11A enhancer dissection by Cas9-mediated in situ saturating mutagenesis. CRISPR-Cas9 gene editing for sickle cell disease and β-thalassemia. The mutational constraint spectrum quantified from variation in 141,456 humans. Insights into human genetic variation and population history from 929 diverse genomes. Variant calling on the GRCh38 assembly with the data from phase three of the 1000 Genomes Project. CRISPRitz: rapid, high-throughput and variant-aware in silico off-target site identification for CRISPR genome editing. Cas-Designer: a web-based tool for choice of CRISPR-Cas9 target sites. CHOPCHOP v3: expanding the CRISPR web toolbox beyond genome editing. Prediction of off-target activities for the end-to-end design of CRISPR guide RNAs. CRISPOR: intuitive guide selection for CRISPR/Cas9 genome editing experiments and screens. Implications of human genetic variation in CRISPR-based therapeutic genome editing. Human genetic variation alters CRISPR-Cas9 on- and off-targeting specificity at therapeutically implicated loci. Evaluation of homology-independent CRISPR-Cas9 off-target assessment methods. Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. DNA targeting specificity of RNA-guided Cas9 nucleases. Tools for experimental and computational analyses of off-target editing by programmable nucleases. Technologies and computational analysis strategies for CRISPR applications. Genome editing with CRISPR-Cas nucleases, base editors, transposases and prime editors. We expect that variant-aware off-target assessment will become integral to therapeutic genome editing evaluation and provide a powerful approach for comprehensive off-target nomination.Īnzalone, A. ![]() This report illustrates how genetic variants should be considered as modifiers of gene editing outcomes. We validated that SpCas9 generates strictly allele-specific indels and pericentric inversions in CD34 + hematopoietic stem and progenitor cells (HSPCs), although high-fidelity Cas9 mitigates this off-target. We tested the software with a BCL11A enhancer targeting guide RNA (gRNA) showing promise in clinical trials for sickle cell disease and β-thalassemia and found that the top candidate off-target is produced by an allele common in African-ancestry populations (MAF 4.5%) that introduces a protospacer adjacent motif (PAM) sequence. We developed an efficient tool called CRISPRme that considers single-nucleotide polymorphism (SNP) and indel genetic variants to nominate and prioritize off-target sites. However, standard computational and biochemical methods to predict off-target potential focus on reference genomes. CRISPR gene editing holds great promise to modify DNA sequences in somatic cells to treat disease.
0 Comments
Leave a Reply. |