Transient Gene Expression is an Effective Experimental Tool for the Research into the Fine Mechanisms of Plant Gene Function: Advantages, Limitations, and Solutions

Tyurin, A. A.; Suhorukova, A. V.; Кабардаева, К. В.; Goldenkova-Pavlova, I. V.

doi:10.3390/plants9091187

Cited by 42 publications

(45 citation statements)

References 83 publications

(180 reference statements)

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“…expression (Tyurin et al 2020). The JASPAR2020 TFBS database has 572 profiles that include different classes and families of TFs in plants based on TF DNA-binding preferences, modeled as position weight matrices (Stormo 2013).…”

Section: Discussionmentioning

confidence: 99%

Promoter analysis of the SPATULA (FvSPT) and SPIRAL (FvSPR) genes in the woodland diploid strawberry (Fragaria vesca L.)

et al. 2021

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The aim of this study was to identify transcription factor (TF) binding sites and cis-regulatory elements (CREs) on the promoters of FvSPR1-like2 (SPIRAL) and FvSPT (SPATULA) genes in the woodland diploid strawberry (Fragaria vesca L.). We identified: (1) MYB59, WRKY25 and WRKY8 TFs which play a role in ethylene signaling; (2) ARF family of TFs which play a role in ARF-mediated auxin signaling on the promoter of FvSPR1-like2 gene; (3) ARR family of TFs which play a role in cytokinin signaling; (4) ERF family of TFs which play a role in ethylene signaling on the promoter of FvSPT. This bioinformatic analysis of TFs and CREs may provide a better understanding of the function of genes involved in, and the mechanism underlying, non-climateric ripening during strawberry fruit maturation.

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Section: Discussionmentioning

confidence: 99%

Promoter analysis of the SPATULA (FvSPT) and SPIRAL (FvSPR) genes in the woodland diploid strawberry (Fragaria vesca L.)

et al. 2021

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“…Specific reviews [ 29 , 30 , 31 , 32 , 33 ] are available on transient gene expression in different host organisms. Sheludko [ 29 ] reviewed the biological characteristics of the transient expression process and application for recombinant protein production.…”

Section: Genome Editing Methodsmentioning

confidence: 99%

“…Sheludko [ 29 ] reviewed the biological characteristics of the transient expression process and application for recombinant protein production. Tyurin et al [ 33 ] also reviewed the main and critical steps involved in various methods for transient gene expression in plants. Another recent review by Zlobin et al [ 4 ] elaborated the utilization of Agrobacterium -based floral dip transformation for genome editing.…”

Section: Genome Editing Methodsmentioning

confidence: 99%

“…The transient expression holds promise before implementing the tedious and time-consuming stable expression methods [ 30 , 94 , 123 ]. The transient transformation has several advantages over stable transformation as mentioned below: Exemption of tissue culture [ 33 ] Temporary expression [ 52 ] T-DNA position effect elimination [ 33 ] Simple, quick, economical, and effective [ 109 ] The transient transformation frequency was at least 1000-fold greater than that of stable transformation [ 29 , 95 ] Transient transformation is often versatile, quick, and efficient as compared to stable transformation [ 99 ]. …”

Section: Stable Vs Transient Genetic Transformationmentioning

confidence: 99%

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Agroinfiltration Mediated Scalable Transient Gene Expression in Genome Edited Crop Plants

Kaur

Manchanda

Kalia

et al. 2021

IJMS

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Agrobacterium-mediated transformation is one of the most commonly used genetic transformation method that involves transfer of foreign genes into target plants. Agroinfiltration, an Agrobacterium-based transient approach and the breakthrough discovery of CRISPR/Cas9 holds trending stature to perform targeted and efficient genome editing (GE). The predominant feature of agroinfiltration is the abolishment of Transfer-DNA (T-DNA) integration event to ensure fewer biosafety and regulatory issues besides showcasing the capability to perform transcription and translation efficiently, hence providing a large picture through pilot-scale experiment via transient approach. The direct delivery of recombinant agrobacteria through this approach carrying CRISPR/Cas cassette to knockout the expression of the target gene in the intercellular tissue spaces by physical or vacuum infiltration can simplify the targeted site modification. This review aims to provide information on Agrobacterium-mediated transformation and implementation of agroinfiltration with GE to widen the horizon of targeted genome editing before a stable genome editing approach. This will ease the screening of numerous functions of genes in different plant species with wider applicability in future.

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“…Transient gene expression in plants was used to test the fluorometric assay for lichenase quantification as an efficient technology for analyzing the functional role of regulatory sequences, which is time-and cost-efficient [25]. For this purpose, the basic vector pLAUMe was constructed.…”

Section: Testing Of Fluorometric Assay For Quantification Of Lichenase In Plants Using Transient Expression Technologymentioning

confidence: 99%

A High Throughput Assay of Lichenase Activity with Congo Red Dye in Plants

Tyurin

Suhorukova

Deineko

et al. 2021

Preprint

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Background : To elucidate the functional role of regulatory sequences and contexts identified and predicted during omics studies in the complex mechanisms of gene expression regulation, experimental verification methods in a plant cell are required. For this, the method of transient expression of reporter genes fused with the tested sequences is widely used. Several reporter systems are available that have shown good performance in the studies including the thermostable lichenase Clostridium thermocellum . However, each reporter system has its own limitations with respect to the quantification of the protein product of a reporter gene and, in particular, for the use of high-throughput approaches. Results: In this study, we report the design a high throughput fluorometric method for quantification of thermostable lichenase C. thermocellum using Congo red and further experimental verification of its relevance and efficiency in assessment of the functional role of regulatory sequences in the plant cell. Conclusion: The specific interaction between the dye Congo red and β -D-glucans formed the background for designing a high-throughput fluorometric assay for quantification of C. thermocellum thermostable lichenase as a reporter protein for plants. This assay (i) makes it possible to precisely measure the amount of reporter protein in a plant sample; (ii) has shown a high sensitivity for quantification of thermostable lichenase; (iii) is more time- and cost-efficient as compared with the Somogyi–Nelson assay; and (iv) is to the least degree dependent on the presence of the tested buffer components as compared with the Somogyi–Nelson assay.

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Transient Gene Expression is an Effective Experimental Tool for the Research into the Fine Mechanisms of Plant Gene Function: Advantages, Limitations, and Solutions

Cited by 42 publications

References 83 publications

Promoter analysis of the SPATULA (FvSPT) and SPIRAL (FvSPR) genes in the woodland diploid strawberry (Fragaria vesca L.)

Promoter analysis of the SPATULA (FvSPT) and SPIRAL (FvSPR) genes in the woodland diploid strawberry (Fragaria vesca L.)

Agroinfiltration Mediated Scalable Transient Gene Expression in Genome Edited Crop Plants

A High Throughput Assay of Lichenase Activity with Congo Red Dye in Plants

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