Aavprime aav system

The plasmid expressing circFoxp1 was synthesized by Sangon (Shanghai, China). The target sequence of circFoxp1 small interfering RNA (siRNA) (siRNA sequence 1#, 5′-TTTTCCCTTTCCAAGGGCACAG-3′; siRNA sequence 2#, 5′-TGACACGGGAACTTTAGAAATGATT-3′) was synthesized by Sangon. The plasmids were packaged into adenoviruses using the AAVPrime AAV System (GeneCopoeia, Inc., Rockville, MD, USA) according to the manufacturer's protocol. The cells were infected with viral at multiplicity of infection = 50 for 48 hours.

The circRNA_101237 vector was synthesized by Sangon Biotech Co., Ltd. circRNA_101237 sequence (1 kb upstream) was inserted into pcDNA3.1. circRNA_101237 without the downstream reverse sequence was used as a negative control. All vectors, including circRNA_101237 and IGF2BP3, were finally cloned into the Adeno-X™ Tet-On Expression System (Seebio Biotech, (Shanghai) Co., Ltd.) according to the manufacturer's protocol. The target sequence of IGF2BP3 small interfering RNA (siRNA) was 5′-AAA TGA TAT TGC TTC CAT GAA TC TT-3′. The target sequence of circRNA_101237 siRNA was 5′-TCT GAT AGT AAG TCT TCG-3′. These siRNA adenoviruses were constructed using the AAVPrime™ AAV System (GeneCopoeia, Inc.) according to the manufacturer's protocol. All constructs were amplified in GCI-AAV-293Ta cells (GeneCopoeia, Inc.). Primary cardiomyocytes were infected with viral at multiplicity of infection=50 for 48 h.