CRISPR-Based Adenine Editors Correct Nonsense Mutations in a Cystic Fibrosis Organoid Biobank

Geurts, Maarten H.; Poel, Eyleen de; Amatngalim, Gimano D.; Oka, Rurika; Meijers, Fleur Michelle; Kruisselbrink, Evelien; Mourik, Peter van; Berkers, Gitte; Groot, K.M. de Winter-de; Michel, Sabine; Muilwijk, Danya; Aalbers, Bente L.; Mullenders, Jasper; Boj, Sylvia F.; Suen, S.W.F.; Brunsveld, Jesse E.; Janssens, Hettie M.; Mall, Marcus; Graeber, Simon Y.; Boxtel, Ruben van; Ent, Cornelis K. van der; Beekman, Jeffrey M.; Clevers, Hans

doi:10.1016/j.stem.2020.01.019

Cited by 160 publications

(129 citation statements)

References 41 publications

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“…However, due to that the incorporation of SV40 disrupts the expression of one of the CFTR alleles 28 , the effect of 5moU-6.3 on correcting CF phenotype (protein expression and electrophysiological function) is likely underestimated in our study. A recent study has used base editor to correct CFTR nonsense mutations in organoid of cystic fibrosis patients 36 . Future work should apply base editing to correct primary CF cells or animal models.…”

Section: Discussionmentioning

confidence: 99%

Chemical modifications of adenine base editor mRNA and guide RNA expand its application scope

et al. 2020

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CRISPR-Cas9-associated base editing is a promising tool to correct pathogenic single nucleotide mutations in research or therapeutic settings. Efficient base editing requires cellular exposure to levels of base editors that can be difficult to attain in hard-to-transfect cells or in vivo. Here we engineer a chemically modified mRNA-encoded adenine base editor that mediates robust editing at various cellular genomic sites together with moderately modified guide RNA, and show its therapeutic potential in correcting pathogenic single nucleotide mutations in cell and animal models of diseases. The optimized chemical modifications of adenine base editor mRNA and guide RNA expand the applicability of CRISPR-associated gene editing tools in vitro and in vivo.

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Section: Discussionmentioning

confidence: 99%

Chemical modifications of adenine base editor mRNA and guide RNA expand its application scope

et al. 2020

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“…in vivo orthotropic imaging, introduction of resistance, target validation, etc. A number of methods have proven useful in delivering gene editing into organoids, electroporation, lentiviral transduction and CRISPR [53][54][55] . Furthermore, genome-wide screens using RNAi-or sgRNA libraries 56 could be powerful tools for discovery of new drug targets.…”

Section: Discussionmentioning

confidence: 99%

A living biobank of matched pairs of patient-derived xenografts and organoids for cancer pharmacology

Xu¹,

Wang²,

Zhou³

et al. 2020

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Patient-derived tumor xenograft (PDX)/organoid (PDO), driven by cancer stem cells (CSC), are considered the most predictive models for translational oncology. Large PDX collections reflective of patient populations have been created and used extensively to test various investigational therapies, including in population-trials as surrogate subjects in vivo. PDOs are recognized as in vitro surrogates for patients amenable for high-throughput screening (HTS). We have built a biobank of carcinoma PDX-derived organoids (PDXOs) by converting an existing PDX library and confirmed high degree of similarities between PDXOs and parental PDXs in genomics, histopathology and pharmacology, suggesting “biological equivalence or interchangeability” between the two. Here we demonstrate the applications of PDXO biobank for HTS “matrix” screening for both lead compounds and indications, immune cell co-cultures for immune-therapies and engineering enables in vitro/in vivo imaging. This large biobank of matched pairs of PDXs/PDXOs across different cancers could become powerful tools for the future cancer drug discovery.

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“…65,66 Adenine base editors have been used to correct nonsense mutations in a CF patient-derived organoid biobank that overrepresents rare CF mutations. 67 In this example, authors used spCas9-and xCas9-derived adenine base editors to correct the R553X and R785X mutations. In another example, researchers used adenosine deaminases to correct the W496X CF mutation, which leads to a premature stop codon.…”

Section: Rna Therapies Can Be Used To Replace or Edit Cftrmentioning

confidence: 99%

Treating Cystic Fibrosis with mRNA and CRISPR

et al. 2020

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Less than 20% of the protein coding genome is thought to be targetable using small molecules. mRNA therapies are not limited in the same way since in theory, they can silence or edit any gene by encoding CRISPR nucleases, or alternatively, produce any missing protein. Yet not all mRNA therapies are equally likely to succeed. Over the past several years, an increasing number of clinical trials with siRNA-and antisense oligonucleotide-based drugs have revealed three key concepts that will likely extend to mRNA therapies delivered by nonviral systems. First, scientists have come to understand that some genes make better targets for RNA therapies than others. Second, scientists have learned that the type and position of chemical modifications made to an RNA drug can alter its therapeutic window, toxicity, and bioavailability. Third, scientists have found that safe and targeted drug delivery vehicles are required to ferry mRNA therapies into diseased cells. In this study, we apply these learnings to cystic fibrosis (CF). We also describe lessons learned from a subset of CF gene therapies that have already been tested in patients. Finally, we highlight the scientific advances that are still required for nonviral mRNA-or CRISPR-based drugs to treat CF successfully in patients.

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CRISPR-Based Adenine Editors Correct Nonsense Mutations in a Cystic Fibrosis Organoid Biobank

Cited by 160 publications

References 41 publications

Chemical modifications of adenine base editor mRNA and guide RNA expand its application scope

Chemical modifications of adenine base editor mRNA and guide RNA expand its application scope

A living biobank of matched pairs of patient-derived xenografts and organoids for cancer pharmacology

Treating Cystic Fibrosis with mRNA and CRISPR

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