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
Citations
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CRISPR screen in mechanism and target discovery for cancer immunotherapy.
TL;DR: An overview of recent progresses in the development of CRISPR-based screens for IO target identification is presented and the challenges and possible solutions in this rapidly growing field are discussed.
Journal ArticleDOI
Precise base editing for the in vivo study of developmental signaling and human pathologies in zebrafish.
Marion Rosello,Marion Rosello,Juliette Vougny,François Czarny,Marina Mione,Jean-Paul Concordet,Shahad Albadri,Filippo Del Bene,Filippo Del Bene +8 more
TL;DR: Zebrafish base editors can generate C-to-T point mutations with high efficiencies without other unwanted on-target mutations as discussed by the authors, which can be used to generate precise point mutations at high efficiency.
Journal ArticleDOI
Designing and executing prime editing experiments in mammalian cells
TL;DR: In this article , the authors provide guidelines for selecting the proper PE system for a given application and how to perform PE in mammalian cells, as well as the design and optimization of pegRNAs.
Journal ArticleDOI
Advances in application of genome editing in tomato and recent development of genome editing technology.
TL;DR: In this article, a review of the application of genome editing in tomato and recent development of the genome editing technology is summarized, and their leaving important enlightenment to plant research and precision plant breeding is also discussed.
Journal ArticleDOI
Synthetic Biology and Tissue Engineering: Toward Fabrication of Complex and Smart Cellular Constructs
Tyler Hoffman,Tyler Hoffman,Petar Antovski,Peyton Tebon,Chun Xu,Chun Xu,Nureddin Ashammakhi,Samad Ahadian,Leonardo Morsut,Ali Khademhosseini +9 more
TL;DR: It is believed that this research area still needs further exploration to fully exploit synthetic biology to make smart and functional cellular constructs for therapeutic and in vitro applications.
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Matthew E. Ritchie,Belinda Phipson,Di Wu,Yifang Hu,Charity W. Law,Wei Shi,Gordon K. Smyth,Gordon K. Smyth +7 more
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Le Cong,Le Cong,F. Ann Ran,F. Ann Ran,David M. Cox,David M. Cox,Shuailiang Lin,Shuailiang Lin,Robert P. J. Barretto,Naomi Habib,Patrick D. Hsu,Patrick D. Hsu,Xuebing Wu,Wenyan Jiang,Luciano A. Marraffini,Feng Zhang +15 more
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Multiplex Genome Engineering Using CRISPR/Cas Systems
Le Cong,F. A. Ran,David Benjamin Turitz Cox,Shuailiang Lin,Robert P. J. Barretto,Naomi Habib,Patrick D. Hsu,Xuebing Wu,Wenyan Jiang,Luciano A. Marraffini,Feng Zhang +10 more
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