TadA reprogramming to generate potent miniature base editors with high precision

share:

Brief intro:

  • Author: Shuqian Zhang, Liting Song, Bo Yuan, Cheng Zhang, Jixin Cao, Jinlong Chen, Jiayi Qiu, Yilin Tai, Jingqi Chen,Zilong Qiu,Xing-Ming Zhao, and Tian-Lin Cheng
  • Journal: Nat Commun
  • Doi: https://www.doi.org/10.1038/s41467-023-36004-2
  • Publication Date: 2023 Jan 26

Products/Services used in the paper

Quotation shows PackGene:5 μl of AAV(1E + 13GC/ml, from PackGene Biotech) were mixed with 5% fast green into the injection syringe. Injection sites at 2/5 of the distance from the lambda suture to each eye were selected for AAV intraventricular injection.

Research Field:miniature CRISPR-Cas12f systems

AAV Serotype:AAV9

Targeted organ:brain

Animal or cell line strain:The Postnatal day 0 (P0) C57BL/6 Mus musculus (Vital River Laboratories) was used in this study.

Request Quote

Abstract

Although miniature CRISPR-Cas12f systems were recently developed, the editing efficacy and targeting range of derived miniature cytosine and adenine base editors (miniCBEs and miniABEs) have not been comprehensively addressed. Moreover, functional miniCBEs have not yet be established. Here we generate various Cas12f-derived miniCBEs and miniABEs with improved editing activities and diversified targeting scopes. We reveal that miniCBEs generated with traditional cytidine deaminases exhibit wide editing windows and high off-targeting effects. To improve the editing signatures of classical CBEs and derived miniCBEs, we engineer TadA deaminase with mutagenesis screening to generate potent miniCBEs with high precision and minimized off-target effects. We show that newly designed miniCBEs and miniABEs are able to correct pathogenic mutations in cell lines and introduce genetic mutations efficiently via adeno-associated virus delivery in the brain in vivo. Together, this study provides alternative strategies for CBE development, expands the toolkits of miniCBEs and miniABEs and offers promising therapeutic tools for clinical applications.

About PackGene

PackGene Biotech is a world-leading CRO and CDMO, excelling in AAV vectors, mRNA, plasmid DNA, and lentiviral vector solutions. Our comprehensive offerings span from vector design and construction to AAV, lentivirus, and mRNA services. With a sharp focus on early-stage drug discovery, preclinical development, and cell and gene therapy trials, we deliver cost-effective, dependable, and scalable production solutions. Leveraging our groundbreaking π-alpha 293 AAV high-yield platform, we amplify AAV production by up to 10-fold, yielding up to 1e+17vg per batch to meet diverse commercial and clinical project needs. Moreover, our tailored mRNA and LNP products and services cater to every stage of drug and vaccine development, from research to GMP production, providing a seamless, end-to-end solution.

Download