Delivering the Goods for Genome Engineering and Editing

作者：Skipper Kristian Al**jerg; Mikkelsen Jacob Giehm^*

来源：Human Gene Therapy, 2015, 26(8): 486-497.

DOI：10.1089/hum.2015.063

A basic understanding of genome evolution and the life and impact of microorganisms, like viruses and bacteria, has been fundamental in the quest for efficient genetic therapies. The expanding tool box for genetic engineering now contains transposases, recombinases, and nucleases, all created from naturally occurring genome-modifying proteins. Whereas conventional gene therapies have sought to establish sustained expression of therapeutic genes, genomic tools are needed only in a short time window and should be delivered to cells ideally in a balanced hit-and-run fashion. Current state-of-the-art delivery strategies are based on intracellular production of protein from transfected plasmid DNA or in vitro-transcribed RNA, or from transduced viral templates. Here, we discuss advantages and challenges of intracellular production strategies and describe emerging approaches based on the direct delivery of protein either by transfer of recombinant protein or by lentiviral protein transduction. With focus on adapting viruses for protein delivery, we describe the concept of all-in-one lentiviral particles engineered to codeliver effector proteins and donor sequences for DNA transposition or homologous recombination. With optimized delivery methodsbased on transferring DNA, RNA, or proteinit is no longer far-fetched that researchers in the field will indeed deliver the goods for somatic gene therapies.

• 出版日期2015-8-1