Search-and-replace genome editing without double-strand breaks or donor DNA
Andrew V. Anzalone,Andrew V. Anzalone,Andrew V. Anzalone,Peyton B. Randolph,Peyton B. Randolph,Peyton B. Randolph,Jessie Rose Davis,Jessie Rose Davis,Jessie Rose Davis,Alexander A. Sousa,Alexander A. Sousa,Alexander A. Sousa,Luke W. Koblan,Luke W. Koblan,Luke W. Koblan,Jonathan M. Levy,Jonathan M. Levy,Jonathan M. Levy,Peter J. Chen,Peter J. Chen,Peter J. Chen,Christine D. Wilson,Christine D. Wilson,Christine D. Wilson,Gregory A. Newby,Gregory A. Newby,Gregory A. Newby,Aditya Raguram,Aditya Raguram,Aditya Raguram,David R. Liu,David R. Liu,David R. Liu +32 more
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TLDR
A new DNA-editing technique called prime editing offers improved versatility and efficiency with reduced byproducts compared with existing techniques, and shows potential for correcting disease-associated mutations.Abstract:
Most genetic variants that contribute to disease1 are challenging to correct efficiently and without excess byproducts2-5. Here we describe prime editing, a versatile and precise genome editing method that directly writes new genetic information into a specified DNA site using a catalytically impaired Cas9 endonuclease fused to an engineered reverse transcriptase, programmed with a prime editing guide RNA (pegRNA) that both specifies the target site and encodes the desired edit. We performed more than 175 edits in human cells, including targeted insertions, deletions, and all 12 types of point mutation, without requiring double-strand breaks or donor DNA templates. We used prime editing in human cells to correct, efficiently and with few byproducts, the primary genetic causes of sickle cell disease (requiring a transversion in HBB) and Tay-Sachs disease (requiring a deletion in HEXA); to install a protective transversion in PRNP; and to insert various tags and epitopes precisely into target loci. Four human cell lines and primary post-mitotic mouse cortical neurons support prime editing with varying efficiencies. Prime editing shows higher or similar efficiency and fewer byproducts than homology-directed repair, has complementary strengths and weaknesses compared to base editing, and induces much lower off-target editing than Cas9 nuclease at known Cas9 off-target sites. Prime editing substantially expands the scope and capabilities of genome editing, and in principle could correct up to 89% of known genetic variants associated with human diseases.read more
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Journal ArticleDOI
CRISPAltRations: a validated cloud-based approach for interrogation of double-strand break repair mediated by CRISPR genome editing.
Gavin Kurgan,Rolf Turk,Heng Li,Nathan Roberts,Garrett R. Rettig,Ashley M. Jacobi,Lauren Tso,Morgan Sturgeon,Massimo Mertens,Roel Noten,Kurt Florus,Mark A. Behlke,Yu Wang,Matthew S. McNeill +13 more
TL;DR: This work characterized DNA repair fingerprints that result from non-homologous end joining after double-stranded breaks were introduced by Cas9 or Cas12a for >500 paired treatment/control experiments and found that building biological understanding of the repair into a novel analysis tool (CRISPAltRations) improved the quality of the results.
Journal ArticleDOI
Nanomedicine for Gene Delivery and Drug Repurposing in the Treatment of Muscular Dystrophies.
Ilaria Andreana,Mathieu Repellin,Mathieu Repellin,Flavia Carton,David Kryza,Stéphanie Briançon,Bénédicte Chazaud,Rémi Mounier,Silvia Arpicco,Manuela Malatesta,Barbara Stella,Giovanna Lollo +11 more
TL;DR: In this article, a review focusing on nanomedicine approaches able to encapsulate therapeutic agents such as small chemical molecules and oligonucleotides to target the most common MDs such as Duchenne Muscular Dystrophy and the Myotonic Dystrophies is provided.
Book ChapterDOI
Globally Important Wheat Diseases: Status, Challenges, Breeding and Genomic Tools to Enhance Resistance Durability
Sridhar Bhavani,Pawan K. Singh,Naeela Qureshi,Xinyao He,Akshaya Kumar Biswal,Philomin Juliana,Abdelfattah A. Dababat,Amira M. I. Mourad +7 more
TL;DR: In this article, an overview of important diseases affecting wheat productivity, considering their geographical distribution, impacts, management strategies and briefly addresses the new molecular/genomic tools in the current era to enhance resistance breeding and deployment opportunities for wheat improvement.
Journal ArticleDOI
Preclinical model systems of ryanodine receptor 1-related myopathies and malignant hyperthermia: a comprehensive scoping review of works published 1990-2019.
TL;DR: Findings from these studies have set the foundation for therapeutic development for MH and RYR1 -RM, a spectrum of rare neuromuscular disorders, which have been developed to better understand underlying pathomechanisms and test potential therapeutics.
Journal ArticleDOI
In vivo hypermutation and continuous evolution
Rosana S. Molina,Gordon Rix,Amanuella Mengiste,Beatriz Alvarez,Daeje Seo,Haiqi Chen,Juan E. Hurtado,Qiong Zhang,Jorge D. García-García,Zachary J. Heins,Patrick J. Almhjell,Frances H. Arnold,Ahmad S. Khalil,Andrew D. Hanson,John E. Dueber,David V. Schaffer,Fei Chen,Seok-Yong Kim,Luis Ángel Fernández,Matthew D. Shoulders,Chang C. Liu +20 more
TL;DR: This work demonstrates the use of OrthoRep to drive the rapid evolution of custom antibodies displayed on the surface of yeast cells, including nanomolar-affinity nanobodies that bind and neutralize SARS-CoV-2.
References
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Journal ArticleDOI
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