Gene editing for cell engineering: trends and applications

Abstract

Gene editing with all its own advantages in molecular biology applications has made easy manipulation of various production hosts with the discovery and implementation of modern gene editing tools such as Crispr (Clustered regularly interspaced short palindromic repeats), TALENs (Transcription activator-like effector nucleases) and ZFNs (Zinc finger nucleases). With the advent of these modern tools, it is now possible to manipulate the genome of industrial production hosts such as yeast and mammalian cells which allows developing a potential and cost effective recombinant therapeutic protein. These tools also allow single editing to multiple genes for knocking-in or knocking-out of a host genome quickly in an efficient manner. A recent study on "multiplexed" gene editing revolutionized the knock-out and knock-in events of yeast and CHO, mammalian cells genome for metabolic engineering as well as high, stable, and consistent expression of a transgene encoding complex therapeutic protein such as monoclonal antibody. The gene of interest can either be integrated or deleted at single or multiple loci depending on the strategy and production requirement. This review will give a gist of all the modern tools with a brief description and advances in genetic manipulation using three major tools being implemented for the modification of such hosts with the emphasis on the use of Crispr-Cas9 for the "multiplexing gene-editing approach" for genetic manipulation of yeast and CHO mammalian hosts that ultimately leads to a fast track product development with consistent, improved product yield, quality, and thus affordability for a population at large.

<p>Cas9 Mediated Correction of β-<em>catenin</em> Mutation and Restoring the Expression of Protein Phosphorylation in Colon Cancer HCT-116 Cells Decrease Cell Proliferation in vitro and Hamper Tumor Growth in Mice in vivo</p>

“…Then, correcting β-catenin genetic mutation by gene-editing technology may develop a next-generation therapeutic approach for colon cancer. [43][44][45] The CRISPR/Cas9 is a prokaryotic nucleic acid-based adaptive immune system to confer resistance to foreign genetic elements such as viral DNA or other foreign DNA. 46,47 Due to the guidance of sgRNA complementary to the target DNA sequence, Cas9 protein binds to a specific genomic locus and creates a site-specific DSB.…”

Sophisticated Cloning, Fermentation, and Purification Technologies for an Enhanced Therapeutic Protein Production: A Review

“…The gene editing tools, CRISPR/Cas9, TALENs, and ZFNs are most commonly used for the host engineering ( Figure 3 ), which may result developing a stable and high producer clone with consistent product quality (Gupta and Shukla, 2016b). Furthermore, glycoengineering is another approach adopted for the production of desired glycoform and good quality product for improved potency.…”

BAPC-assistedCRISPR/Cas9 System: Targeted Delivery into Adult Ovaries for Heritable Germline Gene Editing (Arthropoda: Hemiptera)

“…The relative ease of use, normally by injection into eggs/embryos, combined with improved methods for increase efficacy in CRISPR/Cas9 systems has led CRISPR/Cas to become the primary gene editing tool in the life sciences. For reviews on CRISPR/Cas9 systems see: (Boettcher & McManus 2015; Dominguez et al 2016; Gupta & Shukla 2016; La Russa & Qi 2015; Liang et al 2015; Taning et al, 2017; Wang et al 2016; Wilson & Doudna 2013). Reviews on other gene editing systems with Zinc finger nucleases and TAL effector nucleases, TALENS, see: (Gaj et al, 2013; Bortesi & Fischer 2015; Markert et al, 2016).…”