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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Nuclear dynamics and stress responses in Alzheimer's disease.
TL;DR: In response to extracellular and intracellular stressors, the nucleus and nuclear compartments undergo distinct molecular changes to maintain cell homeostasis as discussed by the authors, in which misfolded proteins and various cellular stressors lead to profound structural and molecular changes at the nucleus.
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Epigenetic marks for mitigating abiotic stresses in plants.
Shahid Ali,Naeem Khan,Yulin Tang +2 more
TL;DR: The functional relevance of epigenetic marks in regulating stress tolerance has been revealed, and epigenetic changes impact the effector genes as discussed by the authors , and plant breeders will benefit from a thorough understanding of these processes to create alternative crop improvement approaches.
Prime Editing for Inherited Retinal Diseases
TL;DR: A new technique known as prime editing (PE) applies a CRISPR-based technology that possesses the potential to correct all twelve possible transition and transversion mutations as well as small insertions and deletions as discussed by the authors.
Journal ArticleDOI
Highly efficient CRISPR-SaKKH tools for plant multiplex cytosine base editing
TL;DR: An effective multiplex cytosine base editor (SaKKHn-pBE) was developed and showed that it recognized NNARRT, NNCR RT, NNGRGT, and NNTRGT PAMs and improved the editing efficiency by modification of the SaKKH sgRNA.
Journal ArticleDOI
Lupus enhancer risk variant causes dysregulation of IRF8 through cooperative lncRNA and DNA methylation machinery
Tian Zhou,Xinyi Zhu,Zhi-Zhong Ye,Yong-Fei Wang,Chao Yao,Ningkun Xu,Mingyan Zhou,Jian yang Ma,Yuting Qin,Yiwei Shen,Yuanjia Tang,Z. Yin,Hong Xu,Yutong Zhang,Xiaoli Zang,Huihua Ding,Wanling Yang,Yanzhi Guo,John B. Harley,Bahram Namjou,Kenneth M. Kaufman,Leah C. Kottyan,Matthew T. Weirauch,Guo Jun Hou,Nan Shen +24 more
TL;DR: In this paper , the locus containing rs2280381 is a cell-type-specific enhancer for IRF8 that spatially interacts with the IRF 8 promoter.
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Multiplex Genome Engineering Using CRISPR/Cas Systems
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