1. Development and Characterization of a Modular CRISPR and RNA Aptamer Mediated Base Editing System
- Author
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Victor M. Tan, Shengkan Jin, Katarzyna M. Tyc, Melany Ruiz-Urigüen, Jennifer A. Harbottle, Ceri M. Wiggins, Juan Collantes, Jinchuan Xing, John J. Lambourne, Hanlin Tao, Chi Su, Amer Alasadi, Jingjing Guo, Huiting Xu, Tommaso Selmi, and Jesse Stombaugh
- Subjects
Computer science ,Base pair ,Green Fluorescent Proteins ,Computational biology ,chemistry.chemical_compound ,INDEL Mutation ,Genome editing ,Exome Sequencing ,Genetics ,Animals ,Humans ,CRISPR ,Clustered Regularly Interspaced Short Palindromic Repeats ,Guide RNA ,Gene ,Research Articles ,Gene Editing ,Bacteria ,Recombinational DNA Repair ,RNA ,Aptamers, Nucleotide ,HEK293 Cells ,chemistry ,Human genome ,RNA Editing ,CRISPR-Cas Systems ,DNA ,RNA, Guide, Kinetoplastida ,Biotechnology - Abstract
Conventional CRISPR approaches for precision genome editing rely on the introduction of DNA double-strand breaks (DSB) and activation of homology-directed repair (HDR), which is inherently genotoxic and inefficient in somatic cells. The development of base editing (BE) systems that edit a target base without requiring generation of DSB or HDR offers an alternative. Here, we describe a novel BE system called Pin-point(TM) that recruits a DNA base-modifying enzyme through an RNA aptamer within the gRNA molecule. Pin-point is capable of efficiently modifying base pairs in the human genome with precision and low on-target indel formation. This system can potentially be applied for correcting pathogenic mutations, installing premature stop codons in pathological genes, and introducing other types of genetic changes for basic research and therapeutic development.
- Published
- 2021