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Mashup Score: 4Taking Gene Therapy to the Next Level: Rahul Kakkar of Tome - 3 month(s) ago
There’s a new genome editing company that everyone is talking about this year. Tome Biosciences came out of stealth in December, claiming the ability to insert DNA sequences of any size at any location across in vivo and ex vivo modalities. Their website says they’re “taking us into the final chapter of medicine.”Tome’s CEO, Rahul Kakkar, joins us today on the program.
Source: www.mendelspod.comCategories: General Medicine News, Future of MedicineTweet
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Mashup Score: 8Sickle Cell Salvation: The Victoria Gray Interview - 3 month(s) ago
Victoria Gray was the first patient to receive a CRISPR-based therapy for sickle cell disease through the exa-cel therapy trial.
Source: www.genengnews.comCategories: General Medicine News, Future of MedicineTweet
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Mashup Score: 3CRISPR Momentum in the Clinic and the Field | The CRISPR Journal - 3 month(s) ago
The CRISPR Journal
Source: www.liebertpub.comCategories: General Medicine News, Future of MedicineTweet
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Mashup Score: 7“CRISPR” Mutations: Inaccurate Linguistic Variations and Misrepresentation of the CRISPR Acronym | The CRISPR Journal - 3 month(s) ago
The CRISPR Journal
Source: www.liebertpub.comCategories: General Medicine News, Future of MedicineTweet
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Mashup Score: 9Generation of Human Isogenic Induced Pluripotent Stem Cell Lines with CRISPR Prime Editing | The CRISPR Journal - 3 month(s) ago
We developed an efficient CRISPR prime editing protocol and generated isogenic-induced pluripotent stem cell (iPSC) lines carrying heterozygous or homozygous alleles for putatively causal single nucleotide variants at six type 2 diabetes loci (ABCC8, MTNR1B, TCF7L2, HNF4A, CAMK1D, and GCK). Our two-step sequence-based approach to first identify transfected cell pools with the highest fraction of edited cells significantly reduced the downstream efforts to isolate single clones of edited cells. We found that prime editing can make targeted genetic changes in iPSC and optimization of system components and guide RNA designs that were critical to achieve acceptable efficiency. Systems utilizing PEmax, epegRNA modifications, and MLH1dn provided significant benefit, producing editing efficiencies of 36–73%. Editing success and pegRNA design optimization required for each variant differed depending on the sequence at the target site. With attention to design, prime editing is a promising appr
Source: www.liebertpub.comCategories: General Medicine News, Future of MedicineTweet
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Mashup Score: 6A landmark partnership for CRISPR drug development | Danaher - 4 month(s) ago
In 2012, Jennifer Doudna and Emmanuelle Charpentier discovered a way to use the molecular complex known as CRISPR for gene editing in the lab. The two were awarded the Nobel Prize in Chemistry, a recognition that typically takes decades to bestow, just eight years after the breakthrough. A mere three years more went by before regulators in the United States and United Kingdom approved the first CRISPR-based therapy, known as Casgevy, for us e in humans. It’s a story of astonishing speed. But the continued
Source: www.danaher.comCategories: General Medicine News, Future of MedicineTweet
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Mashup Score: 1Differential Divalent Metal Binding by SpyCas9's RuvC Active Site Contributes to Nonspecific DNA Cleavage | The CRISPR Journal - 5 month(s) ago
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
Source: www.liebertpub.comCategories: General Medicine News, Future of MedicineTweet
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Mashup Score: 6A PCR-Induced Mutagenesis-Restriction Fragment Length Polymorphism Method for the Detection of CRISPR-Induced Indels | The CRISPR Journal - 5 month(s) ago
As CRISPR-based technologies are widely used for knocking out genes in cell lines and organisms, there is a need for the development of reliable, cost-effective, and fast methods that identify fully mutated clones. In this context, we present a novel strategy named PCR-induced mutagenesis-restriction fragment length polymorphism (PIM-RFLP), which is based on the well-documented robustness and simplicity of the classical PCR-RFLP approach. PIM-RFLP allows the assessment of the editing efficiency in pools of edited cells and the effective identification of fully mutated single-cell clones. It is based on the creation by mutagenic PCR of a restriction enzyme degenerate cleavage site in the PCR product of the wild-type allele, which can then be distinguished from the indel-containing alleles following the standard RFLP procedure. PIM-RFLP is highly accessible, can be executed in a single day, and appears to outperform Sanger sequencing deconvolution algorithms in the detection of fully mut
Source: www.liebertpub.comCategories: General Medicine News, Future of MedicineTweet
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Mashup Score: 1CRISPRi-Mediated Treatment of Dominant Rhodopsin-Associated Retinitis Pigmentosa | The CRISPR Journal - 5 month(s) ago
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
Source: www.liebertpub.comCategories: General Medicine News, Future of MedicineTweet
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Mashup Score: 5The Expanding Dissemination and Distribution Patterns of Diverse CRISPR Plasmids by Addgene | The CRISPR Journal - 5 month(s) ago
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 tec
Source: www.liebertpub.comCategories: General Medicine News, Future of MedicineTweet
RT @mendelspod: Taking Gene Therapy to the Next Level: Rahul Kakkar of @Tome_bio https://t.co/Pxk0qL2Zwd #Genomeediting #CRISPR #geno…