유전자교정기술을 이용한 식물의 phenylpropanoid 대사공학기술 개발 동향

최인호,신용욱,이신우,김윤희 경상국립대학교 농업생명과학연구원 2020 농업생명과학연구 Vol.54 No.5

최근 급속하게 발전하고 있는 유전자교정기술은 식물이 생산하는 특정 이차 대사산물의 집적을 유도하기 위한 식물대사공학연구에 아주 유용하게이용되고 있다. 특히 이들 기술 들을 이용하여 만든 유전자교정 작물 중에 일부는 외래 DNA 단편이 잔존 하지 않기 때문에 기존의 유전자변형작물의안전 관리규정에 적용되지 않을 수 있다는 장점이 있다. 따라서 본 리뷰는 phenylpropanoid 대사과정에 의하여 합성되는 다양한 종류의 이차대사산물을 집적시키기 위한 유전자교정 기술의 적용 연구결과 들을 조사하였다. 먼저, phenylpropanoid 생합성 대사과정에 관여하는 다양한 효소를 암호하는 유전자들을 목표로 하여 식물의 종에 따라 특이하게 집적되는 flavonoids, anthocyanin, 수용성 tannins, 로즈마린산 등의 집적을 유도하거나화색을 변경하는 등의 성공적인 연구결과들을 검토하였다. 또한, phenylpropanoid 대사과정의 조절에 최종조절 스위치 역할을 하는 수많은 종류의MYB 전사인자를 암호 하는 유전자를 목표로 하여 CRISPR 유전자교정을 시도한 연구결과들로부터 식물의 이차세포벽 형성에 관여하는 lignin, 물관부, cellulose 등의 생합성 조절 기작을 이해하고 MYB family에 속하는 수많은 종류의 유전자에 대한 개별적인 기능 분석 연구결과들을조사 분석하여, 문제점 및 향후 연구 방향 등을 검토하였다. Recently the number of approved gene-edited crops for commercialization are gradually increased and, it led us to put intensive investment in the development of gene-edited crops to increase international competitiveness. Developed gene-editing technology has been rapidly applied to a plant metabolic engineering for the accumulation of a particular secondary metabolites. The major advantage is the possible exempt from the regulation under the current GMO laws since gene-editing technology is able to create “a trans-gene free crop” in the final products. In this review, we tried to examine the current research output in the field of metabolic engineering mainly on the phenylpropanoid pathway using CRISPR gene-editing technology. Several genes encoding a particular enzymes involved in the phenylpropanoid pathway have been targeted for the accumulation of particular secondary metabloites such as flavonoids, anthocyanin, soluble tannin, rosmarinic acid, etc. In addition, MYB genes, one of the master switching transcription factor in the phenylpropanoid pathway, have been extensively studied for the dissection of each gene function of extremely divergent MYB gene family and the mechanism of secondary cell wall formation with lignin, xylem and cellulose. We also discussed on the current bottlenecks and further research for the practical application of gene-editing technology on the plant metabolic engineering.

Biomolecular engineering for nanobio/bionanotechnology

Nagamune Teruyuki 나노기술연구협의회 2017 Nano Convergence Vol.4 No.9

Biomolecular engineering can be used to purposefully manipulate biomolecules, such as peptides, proteins, nucleic acids and lipids, within the framework of the relations among their structures, functions and properties, as well as their applicability to such areas as developing novel biomaterials, biosensing, bioimaging, and clinical diagnostics and therapeutics. Nanotechnology can also be used to design and tune the sizes, shapes, properties and functionality of nanomaterials. As such, there are considerable overlaps between nanotechnology and biomolecular engineering, in that both are concerned with the structure and behavior of materials on the nanometer scale or smaller. Therefore, in combination with nanotechnology, biomolecular engineering is expected to open up new fields of nanobio/bionanotechnology and to contribute to the development of novel nanobiomaterials, nanobiodevices and nanobiosystems. This review highlights recent studies using engineered biological molecules (e.g., oligonucleotides, peptides, proteins, enzymes, polysaccharides, lipids, biological cofactors and ligands) combined with functional nanomaterials in nanobio/bionanotechnology applications, including therapeutics, diagnostics, biosensing, bioanalysis and biocatalysts. Furthermore, this review focuses on five areas of recent advances in biomolecular engineering: (a) nucleic acid engineering, (b) gene engineering, (c) protein engineering, (d) chemical and enzymatic conjugation technologies, and (e) linker engineering. Precisely engineered nanobiomaterials, nanobiodevices and nanobiosystems are anticipated to emerge as next-generation platforms for bioelectronics, biosensors, biocatalysts, molecular imaging modalities, biological actuators, and biomedical applications.

Gene Therapy for Bone Tissue Engineering

김영동,Prasad Pofali,박태은,비제이,조기현,Sushila Maharjan,Prajakta Dandekar,Ratnesh Jain,최윤재,Rohidas B. Arote,조종수 한국조직공학과 재생의학회 2016 조직공학과 재생의학 Vol.13 No.2

Gene therapy holds a great promise and has been extensively investigated to improve bone formation and regeneration therapies in bone tissue engineering. A variety of osteogenic genes can be delivered by combining different vectors (viral or non-viral), scaffolds and delivery methodologies. Ex vivo & in vivo gene enhanced tissue engineering approaches have led to successful osteogenic differentiation and bone formation. In this article, we review recent advances of gene therapy-based bone tissue engineering discussing strengths and weaknesses of various strategies as well as general overview of gene therapy.

식물에서의 상동재조합을 이용한 효율적인 진타겟팅 시스템

권용익,이효연 한국식물생명공학회 2015 식물생명공학회지 Vol.42 No.3

The plant breeding technology was developed with genetic engineering. Many researchers and breeders have turned from traditional breeding to molecular breeding. Genetically modified organisms (GMO) were developed via molecular breeding technology. Currently, molecular breeding technologies facilitate efficient plant breeding without introducing foreign genes, in virtue by of gene editing technology. Gene targeting (GT) via homologous recombination (HR) is one of the best gene editing methods available to modify specific DNA sequences in genomes. GT utilizes DNA repair pathways. Thus, DNA repair systems are controlled to enhance HR processing. Engineered sequence specific endonucleases were applied to improve GT efficiency. Engineered sequence specific endonucleases like the zinc finger nuclease (ZFN), TAL effector nuclease (TALEN), and CRISPR-Cas9 create DNA double-strand breaks (DSB) that can stimulate HR at a target site. RecQl4, Exo1 and Rad51 are effectors that enhance DSB repair via the HR pathway. This review focuses on recent developments in engineered sequence specific endonucleases and ways to improve the efficiency of GT via HR effectors in plants.

사상균 Acremonium chrysogenum에서 효모의 D-amino acid oxidase 유전자 발현

김도완,김은주,신재영,강용호 영남대학교 자원문제연구소 2000 資源問題硏究 Vol.19 No.-

Filamentous fungus, Acremonium chrysogenum, producine cephalosporin C antibiotic was co-transformed with PNAVD containing D-amino acid oxidase gene and pDH25 con-taining hygromycin B resistant gene. The integration and the e21pressiul of a yeast Damino acid oxidase gene in A. chrysogenum were certified by PCR and chemiluminescent assay, respectively.

Recent challenges and advances in genetically-engineered cell therapy

용석범,정지영,송윤성,김용희 한국약제학회 2018 Journal of Pharmaceutical Investigation Vol.48 No.2

Cells naturally sense and actively response to their environment. Cell-therapy has long been studied and shown therapeutic effects in various diseases. However, several hurdles should be overcome to improve cell-based therapy. Gene deliverymediated cellular modification has shown improvement of cell function by obstacle gene silencing and therapeutic gene expression. Especially, CRISPR/Cas9-mediated genome editing is a very promising method for gene modification. In this review, we describe the recent advances in genetic modification for cell therapy. Stem cells are still promising source of cell therapy due to their self-renewal character and differentiation potential. Immune cells regulate the inflammatory response and immunization, which inspired various cell therapy using immune-regulatory cells. Conclusively, we emphasize the need to develop gene-modification-based cell therapy as potent future treatment.

The Role of High-throughput Transcriptome Analysis in Metabolic Engineering

Jewett, Michael C.,Oliveira, Ana Paula,Patil, Kiran Raosaheb,Nielsen, Jens The Korean Society for Biotechnology and Bioengine 2005 Biotechnology and Bioprocess Engineering Vol.10 No.5

The phenotypic response of a cell results from a well orchestrated web of complex interactions which propagate from the genetic architecture through the metabolic flux network. To rationally design cell factories which carry out specific functional objectives by controlling this hierarchical system is a challenge. Transcriptome analysis, the most mature high-throughput measurement technology, has been readily applied In strain improvement programs in an attempt to Identify genes involved in expressing a given phenotype. Unfortunately, while differentially expressed genes may provide targets for metabolic engineering, phenotypic responses are often not directly linked to transcriptional patterns, This limits the application of genome-wide transcriptional analysis for the design of cell factories. However, improved tools for integrating transcriptional data with other high-throughput measurements and known biological interactions are emerging. These tools hold significant promise for providing the framework to comprehensively dissect the regulatory mechanisms that identify the cellular control mechanisms and lead to more effective strategies to rewire the cellular control elements for metabolic engineering.

Feature Article : Protein and Gene Delivery in Tissue Engineering

( In Yong Kim ),( Jong Hoon Chung ),( Yun Jaie Choi ),( Chong Su Cho ) 한국조직공학과 재생의학회 2008 조직공학과 재생의학 Vol.5 No.4

In regenerative medicine, growing demand for tissues and organs has led to the rapid development of tissue engineering as an alternative. To improve functional scaffold, many biomaterials serve dual purposes; in addition to providing cell support, cutting-edge scaffolds biologically interact with adhering and invading cells and effectively guide cellular growth and development by releasing bioactive proteins and genes. For achieving the ambitious goal of tissue regeneration and replacement, it is more important to understand the basic principles of protein and gene delivery as well as design of the system required by using proper biomaterials. This review covered the requirement in the materials choice, the strategies, the main techniques for producing proteins and genes releasing scaffolds.

Identification of Gene Networks with Time Delayed Regulation Based on Temporal Expression Profiles

Jeong-Rae Kim,Sang-Mok Choo,Hyung-Seok Choi,Kwang-Hyun Cho IEEE 2015 IEEE/ACM transactions on computational biology and Vol.12 No.5

<P>There are fundamental limitations in inferring the functional interaction structure of a gene (regulatory) network only from sequence information such as binding motifs. To overcome such limitations, various approaches have been developed to infer the functional interaction structure from expression profiles. However, most of them have not been so successful due to the experimental limitations and computational complexity. Hence, there is a pressing need to develop a simple but effective methodology that can systematically identify the functional interaction structure of a gene network from time-series expression profiles. In particular, we need to take into account the different time delay effects in gene regulation since they are ubiquitously present. We have considered a new experiment that measures the overall expression changes after a perturbation on a specific gene. Based on this experiment, we have proposed a new inference method that can take account of the time delay induced while the perturbation affects its primary target genes. Specifically, we have developed an algebraic equation from which we can identify the subnetwork structure around the perturbed gene. We have also analyzed the influence of time delay on the inferred network structure. The proposed method is particularly useful for identification of a gene network with small variations in the time delay of gene regulation.</P>

Improving the secretory capacity of Chinese hamster ovary cells by ectopic expression of effector genes: Lessons learned and future directions

Hansen, H.G.,Pristovsek, N.,Kildegaard, H.F.,Lee, G.M. Pergamon Press ; Elsevier Science Ltd 2017 Biotechnology advances Vol.35 No.1

<P>Chinese hamster ovary (CHO) cells are the preferred cell factory for the production of therapeutic glycoproteins. Although efforts primarily within bioprocess optimization have led to increased product titers of recombinant proteins (r-proteins) expressed in CHO cells, post-transcriptional bottlenecks in the biosynthetic pathway of r-proteins remain to be solved. To this end, the ectopic expression of transgenes (effector genes) offers great engineering potential. However, studies on effector genes have in some cases led to inconsistent results. Whereas this can in part be attributed to product specificity, other experimental and cellular factors are likely important contributors to these conflicting results. Here, these factors are reviewed and discussed with the objective of guiding future studies on effector genes. (C) 2016 Elsevier Inc. All rights reserved.</P>