CRISPER-RNA Guided Gene Editing and Implications in Endogenous Genes Activation
Scholars International Journal of Biochemistry
Short guide RNAs of catalytically inactive CRISPR associated 9 nuclease uses to express endogenous genes in human cells and bacteria. Chimeric nucleases help in precise and efficient genetic modifications leads to double strand breakage which can be repaired by DNA repair mechanism by homology directed repair and non-homology end joining. Here, we describe that single or multiple gRNAs direct dCas9 fused to a VP64 transcriptional activation domain to enhance the expression of endogenous human
... endogenous human genes. Light-activated CRISPR-Cas9 effector (LACE) system induces dynamic regulation of genes in the presence of blue light. Targeted activation of endogenous neurogenic genes in transgenic mice leads to direct and efficient conversion of astrocytes into functional neurons in vivo. CRISPR-on can activate the endogenous IL1RN, SOX2, and OCT4 genes. Activation of genes was achieved by clusters of 3-4 sgRNAs that bind to the proximal promoters, result in their synergistic action in gene induction. Moreover, it fused with transcriptional activators in endogenous human INS (silenced gene with a fully methylated promoter). Multiplexed gene editing helps in metabolic engineering in yeast and CHO mammalian host. In this review we describe the applications and development of Cas9 for a variety of research or translational applications.