CRISPR Del/Rei: a simple, flexible, and efficient pipeline for scarless genome editing

Kyra L Feuer; Marah H Wahbeh; Christian Yovo; Eman Rabie; Anh-Thu N Lam; Sara Abdollahi; Lindsay J Young; Bailey Rike; Akul Umamageswaran; Dimitrios Avramopoulos

doi:10.1038/s41598-022-16004-w

CRISPR Del/Rei: a simple, flexible, and efficient pipeline for scarless genome editing

Sci Rep. 2022 Jul 13;12(1):11928. doi: 10.1038/s41598-022-16004-w.

Authors

Kyra L Feuer^#^{1

2}, Marah H Wahbeh^#^{1

2}, Christian Yovo¹, Eman Rabie^{3

4}, Anh-Thu N Lam¹, Sara Abdollahi¹, Lindsay J Young¹, Bailey Rike¹, Akul Umamageswaran¹, Dimitrios Avramopoulos⁵

Affiliations

¹ McKusick-Nathans Department of Genetic Medicine, Johns Hopkins School of Medicine, Baltimore, USA.
² Predoctoral Training Program in Human Genetics and Molecular Biology, Johns Hopkins School of Medicine, Baltimore, USA.
³ Medical Molecular Genetics Department, Human Genetics and Genome Research Division, National Research Centre, Cairo, Egypt.
⁴ Biotechnology Program, School of Sciences and Engineering, The American University in Cairo, Cairo, Egypt.
⁵ McKusick-Nathans Department of Genetic Medicine, Johns Hopkins School of Medicine, Baltimore, USA. adimitr1@jhmi.edu.

^# Contributed equally.

Abstract

Scarless genome editing of induced pluripotent stem cells (iPSCs) is crucial for the precise modeling of genetic disease. Here we present CRISPR Del/Rei, a two-step deletion-reinsertion strategy with high editing efficiency and simple PCR-based screening that generates isogenic clones in ~ 2 months. We apply our strategy to edit iPSCs at 3 loci with only rare off target editing.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

CRISPR-Cas Systems / genetics
Clustered Regularly Interspaced Short Palindromic Repeats / genetics
Gene Editing*
Genome, Human
Humans
Induced Pluripotent Stem Cells*

Abstract

Publication types

MeSH terms

Grants and funding