Enabling functional genomics with genome engineering

Abstract

Advances in genome engineering technologies have made the precise control over genome sequence and regulation possible across a variety of disciplines. These tools can expand our understanding of fundamental biological processes and create new opportunities for therapeutic designs. The rapid evolution of these methods has also catalyzed a new era of genomics that includes multiple approaches to functionally characterize and manipulate the regulation of genomic information. Here, we review the recent advances of the most widely adopted genome engineering platforms and their application to functional genomics. This includes engineered zinc finger proteins, TALEs/TALENs, and the CRISPR/Cas9 system as nucleases for genome editing, transcription factors for epigenome editing, and other emerging applications. We also present current and potential future applications of these tools, as well as their current limitations and areas for future advances.

Genome Editing: Innovation in Molecular Biology

“…Targeting efficiency of CRISPERCas9 is more than ZFNs and TALENs [1]. It was reported that targeting efficiency of TALENs and ZFNs in human cells lie from 1% to 50% [29][30][31].…”

“…This innovating molecular technique enables addition, deletion and substitution of bases by incorporating detectable changes in the DNA of an organism [1]. In comparison with classical genetic engineering which is the cleavage and random insertion of a foreign gene or DNA sequence from a different specie to another.…”

A Review of CRISPR-Based Genome Editing: Survival, Evolution and Challenges

“…Although most of the genetic syndromes arise from point mutations, modern techniques for correcting point mutation are not efficient and normally induce random insertions and deletions at a target locus subsequent to cellular response to dsDNA breaks (Hilton and Gersbach 2015). However, the development of "base-editing" technique allows the direct, stable transformation of target DNA base into an alternative in a programmable way, without DNA double strand cleavage or a donor template (Komor et al, 2016).…”

A genome editing primer for the hematologist

“…In addition, targeted nucleases may be used to produce additional sequence-specific genetic and epigenetic outcomes that may be exploited to gain knowledge of genome function and produce desirable alterations. 11 Each of the targeted nucleases described below contains 2 major functional moieties, the first of which is specific DNA recognition. The various platforms differ based on the biochemical nature of the recognition (by protein or RNA), by the modularity of recognition (whether 1, 2, or more components are involved), the size of the recognition domain (and its attendant challenges to cellular delivery), the ease with which the interaction may be engineered to recognize a variety of target sequences, and the specificity of such recognition.…”