Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Target-AID-Mediated Multiplex Base Editing in Porcine Fibroblasts

Version 1 : Received: 21 September 2021 / Approved: 21 September 2021 / Online: 21 September 2021 (12:34:20 CEST)

How to cite: Yum, S.; Jang, G.; Koo, O. Target-AID-Mediated Multiplex Base Editing in Porcine Fibroblasts. Preprints 2021, 2021090362 (doi: 10.20944/preprints202109.0362.v1). Yum, S.; Jang, G.; Koo, O. Target-AID-Mediated Multiplex Base Editing in Porcine Fibroblasts. Preprints 2021, 2021090362 (doi: 10.20944/preprints202109.0362.v1).

Abstract

Multiplex genome editing may induce genotoxicity and chromosomal rearrangements due to double-strand DNA breaks at multiple loci simultaneously induced by programmable nucleases, including CRISPR/Cas9. However, recently developed base-editing systems can directly substitute target sequences without double-strand breaks. Thus, the base-editing system is expected to be a safer method for multiplex genome-editing platforms for livestock. Target-AID is a base editing system composed of PmCDA1, a cytidine deaminase from sea lampreys, fused to Cas9 nickase. It can be used to substitute cytosine for thymine in 3-5 base editing windows, 18 bases upstream of the protospacer-adjacent motif site. In the current study, we demonstrated Target-AID-mediated base editing in porcine cells for the first time. We targeted multiple loci in the porcine genome using the Target-AID system and successfully induced target-specific base substitutions with up to 63.15% efficiency. This system can be used for the further production of various genome-engineered pigs.

Keywords

Target-AID; porcine; pig; base editing; CRISPR/Cas

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