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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Genome editing for plant research and crop improvement.
TL;DR: The state-of-the-art development of CRISPR technologies and their applications in plants, from the initial introduction of random small indel (insertion or deletion) mutations at target genomic loci to precision editing such as base editing, prime editing and gene targeting, are summarized in this paper.
Journal ArticleDOI
Fast-forward breeding for a food-secure world.
Rajeev K. Varshney,Rajeev K. Varshney,Rajeev K. Varshney,Abhishek Bohra,Manish Roorkiwal,Manish Roorkiwal,Rutwik Barmukh,Wallace Cowling,Annapurna Chitikineni,Hon-Ming Lam,Lee T. Hickey,Janine Croser,Philipp E. Bayer,David Edwards,José Crossa,Wolfram Weckwerth,Harvey Millar,Arvind Kumar,Michael W. Bevan,Kadambot H. M. Siddique +19 more
TL;DR: In this paper, the authors proposed an approach to accelerate the availability of beneficial alleles for breeding and research using genome sequencing technologies combined with efficient trait mapping procedures, and enhanced interoperability between different omics and phenotyping platforms.
Journal ArticleDOI
Dual-AAV delivering split prime editor system for in vivo genome editing
Shengyao Zhi,Yuxi Chen,Guanglan Wu,Jinkun Wen,Jinni Wu,Qianyi Liu,Yang Li,Rui Kang,Sihui Hu,Jiahui Wang,Puping Liang,Junjiu Huang +11 more
TL;DR: In this article, a new method to deliver PE to adult tissue, paving the way for in-vivo gene-editing therapy using PE, was proposed, where split-PE was used to correct the majority of known human genetic disease-related mutations.
Journal ArticleDOI
CRISPR technology: A decade of genome editing is only the beginning
Joy Y. Wang,Jennifer A. Doudna +1 more
TL;DR: Wang et al. as mentioned in this paper reviewed the origins and utility of CRISPR-based genome editing, the successes and current limitations of the technology, and where innovation and engineering are needed.
Journal ArticleDOI
The glomerular filtration barrier: a structural target for novel kidney therapies
Ilse Daehn,Jeremy S Duffield +1 more
TL;DR: In this paper, a review of therapies and developments focused on maintaining the integrity of the glomerular filtration barrier (GFB) is presented, along with emerging questions in this evolving field.
References
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limma powers differential expression analyses for RNA-sequencing and microarray studies
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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Journal ArticleDOI
Multiplex Genome Engineering Using CRISPR/Cas Systems
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
TL;DR: Two different type II CRISPR/Cas systems are engineered and it is demonstrated that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells, demonstrating easy programmability and wide applicability of the RNA-guided nuclease technology.