Welker Lab

prime editor activity reporter (PEAR)

Prime editing is a CRISPR/Cas9-based gene editing method that allows the integration of short insertions, deletions and substitutions into the genome. The prime editor activity reporter (PEAR) is a plasmid-based, sensitive, fluorescent reporter system for testing and comparing the activity of prime editors. The advantage of the PEAR reporter system is that it can be used to test editing efficiency independently of the genomic context.

Principle of the prime editor activity reporter (PEAR) assay. (A) Schematic of the PEAR. The mechanism of PEAR is based on the same concept as BEAR, and it contains the same inactive splice site. (B) Optimization of PBS, RT, and complementary DNA strand nicking on the PEAR-GFP plasmid. C, D) Flow cytometry measurements of HEK293T cells transfected with active (positive control) or with inactive PEAR plasmids either along with nCas9 for negative controls or with PE2 vectors (black, gray, or green columns, respectively)

Reference
Simon, Dorottya Anna ; Tálas, András ; Kulcsár, Péter István ; Biczók, Zsuzsanna ; Krausz, Sarah Laura ; Várady, György ; Welker, Ervin
PEAR, a flexible fluorescent reporter for the identification and enrichment of successfully primed edited cells
ELIFE 11 Paper: e69504 , 19 p. (2022)

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