ABSTRACT
Gene editing technology is a genetic operation technology that can modify the DNA sequence at the genomic level. The precision gene editing technology based on CRISPR/Cas9 system is a gene editing technology that is easy to operate and widely used. Unlike the traditional CRISPR/Cas9 system, the precision gene editing technology can perform site-directed mutation of genes without DNA template. This review summarizes the recent development of precision gene editing technology based on CRISPR/Cas9, and prospects the challenges and opportunities of this technology.
Subject(s)
Gene Editing , CRISPR-Cas Systems/genetics , Mutation , GenomeABSTRACT
Objective To construct recombinant human Bcl-2 adenoviral vector. Methods Sense human Bcl-2 cDNA fragment from plasimid pcDNA3 1/bcl-2 was cloned into adenovirus shuttle vector pAdCMV. pAdCMV/bcl-2 and plasmid pJM17 were cotransfected into 293 cells with lipofectamine 2000. Replication defective adenovirus was reproduced and purified. Adenoviral virus DNA was detected with polymerase chain reaction(PCR), and plaque assay was performed to determine the titer of virus. Results Recombinant adenovirus containing bcl-2 target gene was obtained. The titer of virus was 2 0?10 10 pfu/ml. Conclusion High titer of adenovirus containing target gene was successfully obtained by recombinant adenovirus vector.