Recent advances in CRISPR-based genome editing technology and its applications in cardiovascular research

Abstract

The rapid development of genome editing technology has brought major breakthroughs in the fields of life science and medicine. In recent years, the clustered regularly interspaced short palindromic repeats (CRISPR)-based genome editing toolbox has been greatly expanded, not only with emerging CRISPR-associated protein (Cas) nucleases, but also novel applications through combination with diverse effectors. Recently, transposon-associated programmable RNA-guided genome editing systems have been uncovered, adding myriads of potential new tools to the genome editing toolbox. CRISPR-based genome editing technology has also revolutionized cardiovascular research. Here we first summarize the advances involving newly identified Cas orthologs, engineered variants and novel genome editing systems, and then discuss the applications of the CRISPR-Cas systems in precise genome editing, such as base editing and prime editing. We also highlight recent progress in cardiovascular research using CRISPR-based genome editing technologies, including the generation of genetically modified in vitro and animal models of cardiovascular diseases (CVD) as well as the applications in treating different types of CVD. Finally, the current limitations and future prospects of genome editing technologies are discussed.

Keywords: Base editing; Blood vessel; CRISPR-Cas system; Cardiovascular disease; Gene therapy; Genome editing; Heart; Prime editing; Transposon-associated genome editing.

Fig. 1

Characteristics of novel Cas orthologs and engineered variants. a Representative type I Cas orthologs capable of large-range deletions. b Representative Cas orthologs of miniature sizes. c Engineered Cas variants with diverse protospacer adjacent motif recognition capabilities. d Structure-guided strategies for improving DNA specificity without affecting the on-target cleavage efficiency

Fig. 2

CRISPR-Cas-based DNA base editing tools. a-c Schematic diagrams of CBE (a), ABE (b), and CGBE (c). d Schematic of PE. UGI uracil-DNA glycosylase inhibitor, AID activation-induced cytidine deaminase, UNG uracil-DNA glycosylase, TadA deoxyadenosine deaminases, RT reverse transcriptase, PBS primer binding site, RTT RT template, CBE cytosine base editor, ABE adenine base editor, CGBE C-to-G base editor, PE Prime editor

Fig. 3

CRISPR-Cas-based transposon systems. Schematic of CRISPR-based transposon systems, CAST system (a) and FiCAT system (b), which mediate site-specific DNA integration. Tns Tn7-like transposases, PB piggyBac transposase, LE transposon left end sequences, RE transposon right end sequences, CAST CRISPR-associated transposon, FiCAT find and cut-and-transfer