Metabolic Mechanisms and Diabetes Candidate Genes: Insights Gained from Genetic Engineering with Recombinant Adenoviruses

Author(s):  
H.K. Berman ◽  
K.Y. Trinh ◽  
R.M. O�Doherty ◽  
P.A. Antinozzi ◽  
R. Gasa ◽  
...  
2021 ◽  
Author(s):  
◽  
Christopher Willig

Plant genetic engineering relies on the ability to transmit and express cloned DNA sequences in plant cells (transformation) as well as the capacity for the cells carrying this DNA to undergo division and differentiation (regeneration), eventually giving rise to a mature whole plant. The breadth of application for genetic engineering is limited by constraints on one or both of these factors in many plant species and individual varieties. Uncovering plant genes which are involved in important aspects of either component can inform the development of technologies that serve to enable or improve the efficiency of genetic modification methods. The most commonly employed method of delivering exogenous genetic material into plant cells is via disarmed strains of the plant pathogen Agrobacterium tumefaciens. Somatic embryogenesis is a frequently applied mode of plant regeneration following DNA delivery, especially in major cereal crops such as maize, rice, and sorghum. In the work reported here, whole transcriptome sequencing (RNA-seq) was used in two different experiments to capture transcriptional dynamics throughout early somatic embryogenesis in immature zygotic embryo tissue of the major crop plant sorghum (Sorghum bicolor), and throughout early times following host plant inoculation with A. tumefaciens in seedlings of the model plant Arabidopsis thaliana (Chapters 2 and 3, respectively). In both cases, differential expression analysis revealed many genes which were induced either during somatic embryogenesis or in response to inoculation with either virulent or avirulent A. tumefaciens strains. Several of these genes were highlighted as candidates for future study into their potential role in the respective processes. Multiple candidate genes were functionally tested, using transgenic methods, for the possibility of having a role in the regulation of somatic embryogenesis in sorghum (Chapter 4). These experiments failed to confirm an influence over the process for all candidate genes that were evaluated. The experimental work documented herein contributes to a growing body of literature documenting plant genes which could serve as possible targets for techniques that work to enhance the utility of biotechnological methods for improving traits in plants.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Dandan Li ◽  
Qing Wang ◽  
Xin Xu ◽  
Jingsheng Yu ◽  
Zhiyu Chen ◽  
...  

Abstract Background The investigation of molecular mechanisms involved in lipid metabolism plays a critical role for the genetic engineering of safflower (Carthamus tinctorius L.) to increase the oil accumulation level or to change the oil composition. Although transcript sequences are currently available for the leaves and flowers of safflower, a wide range scan of temporal transcripts at different stages of seed development has not been conducted for safflower. Results In this study, temporal transcriptome sequencing was executed at 10, 14, 18, and 22 days after flowering (DAF) to uncover the molecular networks concerned in the biosynthesis of unsaturated fatty acids (USFAs). The results revealed that the biosynthesis of fatty acids is a dominant cellular process from 10 to 14 DAF, while degradation mainly happens after 18 DAF. Significant expression changes of two genes, stearoyl-[acyl-carrier-protein] 9-desaturase gene (SAD) from 10 to 14 DAF and oleate desaturase (FAD2–1) from 14 to 18 DAF, were detected at the transcriptomic levels, and the temporal expression patterns revealed by the transcriptomic analysis were confirmed using quantitative real-time PCR experiments. In addition, 13 candidate transcription factors (TFs) involved in regulating the expression level of the FAD2–1 gene were identified. Conclusions These results create a link between fatty acid biosynthesis and gene expression at different developmental stages of the seeds, provide insight into the underlying lipid metabolism, and meanwhile lay an important foundation for the genetic engineering of safflower varieties. We have identified novel candidate genes, including TFs, that are worthy of further exploration.


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