Gene editing for cell engineering: trends and applications

被引:79
作者
Gupta, Sanjeev K. [1 ,2 ]
Shukla, Pratyoosh [1 ]
机构
[1] Maharshi Dayanand Univ, Dept Microbiol, Enzyme Technol & Prot Bioinformat Lab, Rohtak, Haryana, India
[2] Ipca Labs Ltd, Ctr Res & Dev, Adv Biotech Lab, Bombay, Maharashtra, India
关键词
Genome editing; Crispr-Cas9; TALENs; ZFNs; multiplexed gene editing; CHO cell; yeast; RECOMBINANT PROTEIN THERAPEUTICS; DOUBLE-STRAND BREAKS; ONE-STEP GENERATION; ZINC-FINGER; CHO-CELLS; DNA-CLEAVAGE; MAMMALIAN-CELLS; GENOME; CRISPR; EFFICIENT;
D O I
10.1080/07388551.2016.1214557
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:672 / 684
页数:13
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