Advances in Genome Engineering Approaches

Arvind Chhabra

doi:10.4172/2169-0111.1000e107

Abstract

Targeted genome editing is essential for functional characterization of a gene of interest. Targeted gene inactivation via homologous recombination made it feasible to create gene knockout animal models to ascertain the physiological role of the target genes; however, lower efficiency of site specific insertion of the genetically modified construct through homologous recombination has limited a wider applicability of this approach. Development of targeted gene knockdown through RNA interferce (RNAi) offered a cost effective, high-throughput alternative to homologous recombination, however, RNAi-mediated gene knockdown is incomplete, produces experiment to experiment variation, and could provide only a temporary inhibition of the gene function. Development of genome engineering methodologies utilizing nucleases linked to the guide sequences targeting a gene of interest, such as Zinc Finger Nucleases (ZFN), Transcription Activator like Effector Nucleases (TALEN) and Clustered Palindromic Repeats (CRISPR), are quite encouraging. A brief overview of recent advances in genome engineering approaches is provided with their respective advantages and limitations.

Highlights

Targeted genome editing is essential for functional characterization of a gene of interest
These double-stranded products assemble with Argonaute proteins in a protein complex known as the RNA-induced silencing complex (RISC) [3,4,5], such that their one strand is preferentially selected to guide the sequence-specific modulation of complementary target mRNA through mRNA cleavage, translational repression, or cleavage independent mRNA decay [3,4,5]
While on one hand these findings demonstrate an active role for microRNAs in modulation of cellular physiology, it offers a unique opportunity for targeted genome engineering

Summary

Introduction

Targeted genome editing is essential for functional characterization of a gene of interest. Identification of naturally occurring microRNAs that can regulate specific physiologic pathways would be quite significant in developing microRNA based genome engineering methodologies.

Results

Conclusion