Bharucha Nike, Ataam Jennifer Arthur, Gavidia Alexandra A, Karakikes Ioannis
Department of Cardiothoracic Surgery and Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.
Department of Cardiothoracic Surgery and Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.
Stem Cell Res. 2021 Dec;57:102610. doi: 10.1016/j.scr.2021.102610. Epub 2021 Nov 24.
Prime editing uses the Cas9 nickase fused to a reverse transcriptase to copy a DNA sequence into a specific locus from a 'prime editing' guide RNA (pegRNA), eliminating the need for double-stranded DNA breaks and donor DNA templates. To facilitate prime editing in human induced pluripotent stem cells (iPSCs), we integrated a doxycycline-inducible Prime Editor protein (PE2) into the AAVS1 genomic safe harbor locus. Prime editing of iPSCs resulted in precise insertion of three nucleotides in HEK3 locus with high efficiency, demonstrating the utility of this approach. This engineered cell line can be used to edit a single or multiple genomic loci by introducing a target-specific pegRNA for precise and effective genome editing to facilitate disease modeling and functional genetics studies.
引导编辑利用与逆转录酶融合的Cas9切口酶，将DNA序列从“引导编辑”引导RNA（pegRNA）复制到特定位点，无需双链DNA断裂和供体DNA模板。为便于在人诱导多能干细胞（iPSC）中进行引导编辑，我们将强力霉素诱导型引导编辑器蛋白（PE2）整合到AAVS1基因组安全港位点。对iPSC进行引导编辑可高效地在HEK3位点精确插入三个核苷酸，证明了该方法的实用性。通过引入针对特定靶点的pegRNA进行精确有效的基因组编辑，这种工程细胞系可用于编辑单个或多个基因组位点，以促进疾病建模和功能遗传学研究。
