Prime Editing: An All-Rounder for Genome Editing

Abstract

Prime editing (PE), as a “search-and-replace” genome editing technology, has shown the attractive potential of versatile genome editing ability, which is, in principle, currently superior to other well-established genome-editing technologies in the all-in-one operation scope. However, essential technological solutions of PE technology, such as the improvement of genome editing efficiency, the inhibition of potential off-targets and intended edits accounting for unexpected side-effects, and the development of effective delivery systems, are necessary to broaden its application. Since the advent of PE, many optimizations have been performed on PE systems to improve their performance, resulting in bright prospects for application in many fields. This review briefly discusses the development of PE technology, including its functional principle, noteworthy barriers restraining its application, current efforts in technical optimization, and its application directions and potential risks. This review may provide a concise and informative insight into the burgeoning field of PE, highlight the exciting prospects for this powerful tool, and provide clues for questions that may propel the field forward.

CRISPR/Cas9 therapeutics: progress and prospects

“…Thus, a new single-base-editing technology may be needed. 169 The prime editor (PE) consists of two parts, a reverse transcriptase (RT) protein from Moloney murine leukemia virus (M-MLV) fused with Cas9n (H840A) and a 30 bp sgRNA (pegRNA), including a primer binding site (PBS) and an RT template. 27,[170][171][172][173] After Cas9n reaches the designated position, it cuts the target DNA strand.…”

Current advancement in the application of prime editing

“…However, comparing the CRISPR-Cas system, the editing efficiency of PE is still very low. Meanwhile, it was reported that the editing efficiency of PE system was various in different species ( Lu et al, 2022 ), which could be related to the expression level of PE effecter protein and pegRNA in different species. Second, the ratio of off-target editing is needed to be reduced and the mechanism of off-target editing is needed to be clarified.…”

“…Recently, several comprehensive reviews on progress of the PE strategies and applications have been published ( Bharucha et al, 2022 ; Chen & Liu, 2022 ; Doman et al, 2022 ; Lu et al, 2022 ; Raguram et al, 2022 ). The PE strategies for genome manipulation and their limitations are summarized.…”

Base and Prime Editing in the Retina—From Preclinical Research toward Human Clinical Trials

“…In addition, enhanced pegRNAs [ 19 ] have been generated that install a structured RNA pseudoknot at the 3′ end to protect it from exonucleases by increasing RNA stability. Overall, prime editing not only maintains the advantages of evading a double stranded break, but also increases versatility and allows for a broader editing range [ 20 ] by being able to install any single base-to-base change, delete at least 80 nucleotides, and insert at least 44 nucleotides [ 18 , 21 ].…”