Transgenic Research最新文献

Shaker K⁺ channel NKT3A enhances potassium uptake and transport in tobacco (Nicotiana tabacum L.) seedlings under low potassium stress.

IF 2.7 3区生物学

Transgenic Research Pub Date : 2025-03-27 DOI: 10.1007/s11248-024-00419-4

Haiying Xiang, Guang Yuan, Chuhan Shi, Li Xu, Jianduo Zhang, Qili Mi, Qian Gao, Wenwu Yang, Haitao Huang, Kunmiao Wang, Wanli Zeng, Yang Ning, Qian Wang

{"title":"Shaker K+ channel NKT3A enhances potassium uptake and transport in tobacco (Nicotiana tabacum L.) seedlings under low potassium stress.","authors":"Haiying Xiang, Guang Yuan, Chuhan Shi, Li Xu, Jianduo Zhang, Qili Mi, Qian Gao, Wenwu Yang, Haitao Huang, Kunmiao Wang, Wanli Zeng, Yang Ning, Qian Wang","doi":"10.1007/s11248-024-00419-4","DOIUrl":"10.1007/s11248-024-00419-4","url":null,"abstract":"One of the nutrients that is necessary for plant growth and development is potassium (K+). The uneven production and distribution of global potassium resources significantly challenge crop yields and quality. A moderate increase in the potassium content within plants can enhance both crop yield and quality. This study identifies the Shaker K+ channel NKT3A within the model crop, tobacco. The yeast heterologous expression system demonstrated its capability for K+ inward transportation. GUS staining and RT-qPCR analyses of the constructed promoter materials revealed NKT3A's activity during the tobacco seedling stage. Expression levels are higher in the leaf and stems, with low potassium levels inducing upregulation of its expression, also observed in roots. Gene editing technology was employed to construct overexpression and knockout mutants, with subsequent measurement of their phenotypes. Results indicate that NKT3A expression enhances facilitates potassium absorption and transport in tobacco seedlings under low potassium conditions. For the first time, this article identifies the Shaker potassium channel gene NKT3A, which functions as an inward rectifier K+ channel in tobacco. It elucidates the gene's role in regulating potassium distribution under low potassium conditions, thereby deepening our understanding of plant responses in such environments and offering a potential target for enhancing crop potassium use efficiency.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"17"},"PeriodicalIF":2.7,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143721573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Plant-derived recombinant macromolecular PAP-IgG Fc as a novel prostate cancer vaccine candidate eliciting robust immune responses.

IF 2.7 3区生物学

Transgenic Research Pub Date : 2025-03-26 DOI: 10.1007/s11248-025-00433-0

Yangjoo Kang, Deuk-Su Kim, Hyunjoo Hwang, Yerin Kim, Young-Jin Seo, Peter Hinterdorfer, Kisung Ko

{"title":"Plant-derived recombinant macromolecular PAP-IgG Fc as a novel prostate cancer vaccine candidate eliciting robust immune responses.","authors":"Yangjoo Kang, Deuk-Su Kim, Hyunjoo Hwang, Yerin Kim, Young-Jin Seo, Peter Hinterdorfer, Kisung Ko","doi":"10.1007/s11248-025-00433-0","DOIUrl":"https://doi.org/10.1007/s11248-025-00433-0","url":null,"abstract":"Prostatic acid phosphatase (PAP) is a specific protein that is highly expressed in prostate cancer. In this study, we constructed two recombinant PAP fusion genes: PAP fused to the immunoglobulin G (IgG) Fc fragment (designated PAP-Fc) and PAP-Fc fused to the endoplasmic reticulum retention sequence KDEL (designated PAP-FcK). Transgenic Nicotiana tabacum plants expressing these recombinant macromolecular proteins (MPs) were generated using Agrobacterium-mediated transformation, and the presence of both genes was confirmed through genomic PCR. Western blot analysis validated the expression of PAP-Fc and PAP-FcK MPs, which were successfully purified via protein A affinity chromatography. Size-exclusion high-performance liquid chromatography revealed dimeric peaks for PAP-Fc (PAP-FcP) and PAP-FcK (PAP-FcKP). Bio-transmission electron microscopy demonstrated 'Y'-shaped protein particles resembling antibody structures. Moreover, PAP-FcP and PAP-FcKP exhibited a high association rate with human FcγR and FcRn. Vaccination of mice with both PAP-FcP and PAP-FcKP resulted in increased total IgG against PAP and enhanced activation of CD4+ T cells, comparable to mice immunized with PAP, which served as a positive control. These findings indicate that both plant-derived MPs can effectively induce adaptive immunity, positioning them as promising candidates for prostate cancer vaccines. Overall, plants expressing PAP-Fc and PAP-FcK represent a viable production system for antigenic macromolecule-based prostate cancer vaccines.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"16"},"PeriodicalIF":2.7,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143721572","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Low-temperature embryo incubation suppresses off-target mutagenesis during CRISPR-Cas9 genome editing in medaka (Oryzias latipes) and zebrafish (Danio rerio).

IF 2.7 3区生物学

Transgenic Research Pub Date : 2025-03-25 DOI: 10.1007/s11248-025-00434-z

Takashi Yamanaka, Akiko Sogo, Shingo Maegawa, Masato Kinoshita

{"title":"Low-temperature embryo incubation suppresses off-target mutagenesis during CRISPR-Cas9 genome editing in medaka (Oryzias latipes) and zebrafish (Danio rerio).","authors":"Takashi Yamanaka, Akiko Sogo, Shingo Maegawa, Masato Kinoshita","doi":"10.1007/s11248-025-00434-z","DOIUrl":"https://doi.org/10.1007/s11248-025-00434-z","url":null,"abstract":"Gene knockout using CRISPR-Cas9 is often employed in research aimed at elucidating gene functions in fish. However, CRISPR-Cas9 sometimes introduces unintended alterations, known as off-target mutations. These mutations can reduce the robustness of data during phenotypic analysis. In this study, we focused on the culture temperature, which is known to significantly influence mutagenesis, and examined whether low-temperature culture after introducing CRISPR-Cas9 into early embryos of medaka and zebrafish suppresses off-target mutations. Continuous incubation of medaka at 16 °C significantly reduced off-target mutation rates compared to those at 28 °C; the drawback is that it decreased the survival rate of medaka embryos. Therefore, low-temperature incubation was limited to early development in both zebrafish and medaka, and then the temperature was increased to 28 °C. Under these conditions, the mutation rates of the three off-target regions in medaka (Off-D, Off-P, and Off-A) significantly decreased, whereas those of the three target regions (DJ-1, p4hb, and avt) were unaffected. Similarly, the mutation rate of the zebrafish target region (ywhaqa) remained high, whereas the off-target (Off-Y1) mutation rate significantly reduced. Furthermore, this method effectively suppressed the germ line transmission of off-target mutations in medaka. This approach is effective to obtain more reliable data from the G0 generation of medaka and zebrafish and may reduce the screening effort required to remove individuals with off-target mutations in the F1 generation.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"15"},"PeriodicalIF":2.7,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143711379","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

An advanced cytosine base editor enabled the generation of cattle with a stop codon in the β-lactoglobulin gene.

IF 2.7 3区生物学

Transgenic Research Pub Date : 2025-03-14 DOI: 10.1007/s11248-025-00435-y

Qiang Ding, Zhaokang Cui, Qianqian Shi, Yan Zhang, Nan He, Rihong Guo, Yu Tian, Shaoxian Cao, Jifeng Zhong, Huili Wang

{"title":"An advanced cytosine base editor enabled the generation of cattle with a stop codon in the β-lactoglobulin gene.","authors":"Qiang Ding, Zhaokang Cui, Qianqian Shi, Yan Zhang, Nan He, Rihong Guo, Yu Tian, Shaoxian Cao, Jifeng Zhong, Huili Wang","doi":"10.1007/s11248-025-00435-y","DOIUrl":"https://doi.org/10.1007/s11248-025-00435-y","url":null,"abstract":"β-Lactoglobulin (BLG) is an allergen present in milk that can induce an acute immune response in certain individuals. The successful use of cytosine base editors (CBEs) can introduce stop codons into premature mRNA, thereby generating animals with disrupted genes that negatively regulate target traits. In this study, we employed a CBE system to target the major milk allergen BLG in bovine embryos, mammary epithelial cells, and live cattle. First, the precise single-base editing of the BLG gene in bovine embryos was achieved by designing an effective sgRNA to induce a c.61C > T substitution in the coding region, converting codon 21Gln (p.21Gln) to a premature stop codon. Sanger sequencing revealed an editing efficiency of 83.3% (20 out of 24 embryos), including two homozygous edits. Second, a bovine mammary epithelial cell line harboring BLG edits was constructed using the same CBE system. Sequencing showed that the designed sgRNA1 enabled the simultaneous conversion of three consecutive cytosines (c.59-61CCC > TTT) to thymines. At position c.61, single-cell clones exhibited monoallelic or biallelic editing (BLGc.61C > T), with monoallelic edits at positions c.59 and c.60 (CC > TT). Gene expression analysis confirmed that the BLGc.61C > T mutation effectively suppressed BLG expression at both the mRNA and protein levels, even in monoallelically edited cells. Finally, we successfully generated a heterozygous BLGc.61C > T single-base-edited dairy cow that despite its heterozygosity, showed significantly reduced BLG expression in the mammary epithelial cells and milk. Collectively, this study demonstrates the feasibility of using CBEs to disrupt BLG expression in dairy cows and provides a foundation for application in generating hypoallergenic dairy products.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"14"},"PeriodicalIF":2.7,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143634684","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Genome editing research initiatives and regulatory landscape of genome edited crops in India.

IF 2.7 3区生物学

Transgenic Research Pub Date : 2025-03-13 DOI: 10.1007/s11248-025-00432-1

Neha Sharma, Kanika Thakur, Rasna Zinta, Vikas Mangal, Dalamu, Jagesh Kumar Tiwari, Salej Sood, Som Dutt, Vinod Kumar, Brajesh Singh, Ajay Kumar Thakur

{"title":"Genome editing research initiatives and regulatory landscape of genome edited crops in India.","authors":"Neha Sharma, Kanika Thakur, Rasna Zinta, Vikas Mangal, Dalamu, Jagesh Kumar Tiwari, Salej Sood, Som Dutt, Vinod Kumar, Brajesh Singh, Ajay Kumar Thakur","doi":"10.1007/s11248-025-00432-1","DOIUrl":"https://doi.org/10.1007/s11248-025-00432-1","url":null,"abstract":"Food and nutritional security are the top priorities in Indian agriculture. Exponential population growth coupled with climate change effects has become a serious challenge for sustainable agriculture. Genome editing has revolutionized the agricultural sector because of its ability to create precise, stable and predictable modifications in the genome and therefore, offers great opportunities for crop improvement in India. However, for harvesting the real benefits of this technology in agriculture sector, there is a strong need of creating awareness among the end users and development of suitable policies for regularization of genome edited products. Many regulatory agencies around the world have been modernizing their regulatory approaches to be more risk proportionate and to reflect a more science-based approach. In this article, recent research initiatives and developments undertaken by different Indian institutes/organizations for the genetic improvement of agricultural and horticultural crops via genome editing technologies are summarized. Furthermore, to benefit from this potential technology in our country, regulatory policies must be clear, science-based and proportionate. Therefore, in the present review, the regulatory policies related to the genome editing of crop products in India are discussed in detail. This review will sensitize researchers and stakeholders to the application of genome editing techniques in crop improvement and various biosafety committees involved in the development and regulation of genome edited crops.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"13"},"PeriodicalIF":2.7,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143626135","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Comprehensive transcriptomic analysis of myostatin-knockout pigs: insights into muscle growth and lipid metabolism.

IF 2.7 3区生物学

Transgenic Research Pub Date : 2025-02-20 DOI: 10.1007/s11248-025-00431-2

Nasar Khan, Zhouyan Li, Akbar Ali, Biaohu Quan, Jindan Kang, Munib Ullah, Xi-Jun Yin, Muhammad Shafiq

{"title":"Comprehensive transcriptomic analysis of myostatin-knockout pigs: insights into muscle growth and lipid metabolism.","authors":"Nasar Khan, Zhouyan Li, Akbar Ali, Biaohu Quan, Jindan Kang, Munib Ullah, Xi-Jun Yin, Muhammad Shafiq","doi":"10.1007/s11248-025-00431-2","DOIUrl":"https://doi.org/10.1007/s11248-025-00431-2","url":null,"abstract":"Pigs are a vital source of protein worldwide, contributing approximately 43% of global meat production. Recent genetic advancements in the myostatin (MSTN) gene have facilitated the development of double-muscling traits in livestock. In this study, we investigate the transcriptomic profiles of second-generation MSTN-knockout (MSTN-/-) pigs, generated through CRISPR/Cas9 gene editing and somatic cell nuclear transfer (SCNT). Using RNA sequencing, we compared the transcriptomic landscapes of muscle tissues from MSTN-/- pigs and wild-type (WT) counterparts. The sequencing yielded an average unique read mapping rate of 86.7% to the Sus scrofa reference genome. Our analysis revealed 15,142 differentially expressed genes (DEGs), including 121 novel genes, with 2554 genes upregulated and 1629 downregulated in the MSTN-/- group relative to the wild-type group. Notable transcriptomic changes were identified in genes associated with muscle development, lipid metabolism, and other physiological processes. These findings provide valuable insights into the molecular consequences of MSTN inactivation, with potential applications in the optimization of livestock breeding and advancements in biomedical research.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"12"},"PeriodicalIF":2.7,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143469350","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Effect of transgene on salt tolerance of tobacco.

IF 2.7 3区生物学

Transgenic Research Pub Date : 2025-01-28 DOI: 10.1007/s11248-025-00430-3

Jie Sun, Yan Dong, Yuemei Meng, Jingran Bi, Hongmei Liu, Junjie Ren, Jinmao Wang, Yachao Ren, Minsheng Yang

{"title":"Effect of transgene on salt tolerance of tobacco.","authors":"Jie Sun, Yan Dong, Yuemei Meng, Jingran Bi, Hongmei Liu, Junjie Ren, Jinmao Wang, Yachao Ren, Minsheng Yang","doi":"10.1007/s11248-025-00430-3","DOIUrl":"10.1007/s11248-025-00430-3","url":null,"abstract":"To explore the effects of salt-tolerance gene accumulation on salt tolerance in transgenic plant, we used four types of plant expression vector (N27, N28, N29, and N30) carrying mtlD, mtlD + gutD, mtlD + gutD + BADH, mtlD + gutD + BADH + sacB genes respectively, to transform tobacco through Agrobacterium-mediated method. Transgenic lines were identified through polymerase chain reaction (PCR) detection. Transgenic lines and non-transgenic plant (CK) were subjected to 6‰ sodium chloride solution stress; then, fluorescence quantitative PCR (FQ-PCR) and salt tolerance indexes were used to assess characteristics. PCR showed the exogenous genes had been integrated into the tobacco genome. FQ-PCR showed under clean water treatment the target genes were expressed in all transgenic plants at the transcriptional level. The transcript abundances of target genes changed with the number of genes increased, and improved following salt stress. Comparative analyses of salt tolerance indexes showed height growth, biomass (except for N29), chlorophyll content, net photosynthetic rate, Fv/Fm, and PI of all transgenic plants and CK were lower under salt stress than under clean water treatment, to varying degrees. However, the descent ratio was smaller in transgenic plants. A comprehensive evaluation of multiple salt-tolerance indicators performed using the membership function method showed the average salt tolerance of each vector transgenic line was higher than that of CK, and salt tolerance was greater in transgenic polyvalent gene lines than in transgenic monovalent gene lines. The average salt tolerance was N29 > N28 > N30 > N27 > CK. This study provides a theoretical and practical reference for salt tolerance breeding in other plants.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"11"},"PeriodicalIF":2.7,"publicationDate":"2025-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143053782","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

NtLPA1 overexpression regulates the growth of tobacco and enhances resistance to blight. NtLPA1过表达调控烟草生长，增强烟草抗枯萎病能力。

IF 2.7 3区生物学

Transgenic Research Pub Date : 2025-01-09 DOI: 10.1007/s11248-024-00420-x

Guiqin Shi, Yanxiao Bu, Lei Chi, Xifeng Zhang, Yuqing Meng, Shijie Zhang, Geng Tian

引用次数: 0

Expression of Agrobacterium Isopentenyl transferase (IPT) gene in wheat improves drought tolerance. 异戊烯基转移酶（IPT）基因在小麦中的表达提高了小麦的抗旱性。

IF 2.7 3区生物学

Transgenic Research Pub Date : 2025-01-09 DOI: 10.1007/s11248-024-00421-w

Sidra Ijaz, Aftab Bashir, Kauser A Malik

{"title":"Expression of Agrobacterium Isopentenyl transferase (IPT) gene in wheat improves drought tolerance.","authors":"Sidra Ijaz, Aftab Bashir, Kauser A Malik","doi":"10.1007/s11248-024-00421-w","DOIUrl":"https://doi.org/10.1007/s11248-024-00421-w","url":null,"abstract":"Drought, as an abiotic stressor, globally limits cereal productivity, leading to early aging of leaves and lower yields. The expression of the isopentenyl transferase (IPT) gene, which is involved in cytokinin (CK) biosynthesis, can delay drought-induced leaf senescence. In this study, the Agrobacterium Isopentenyl transferase (IPT) gene was introduced into two local hexaploid wheat cultivars, NR-421 and FSD-2008. The expression cassette was developed containing the IPT gene under transcriptional regulation of the stress-inducible promoter 'Dehydrin,' sourced from Hordeum vulgare. The gene expression cassette was assembled in pSB219M, a modified transformation vector for monocots, equipped with both an antibiotic (spectinomycin) and an herbicide selection marker (BASTA). Initial screening of transgenic plants involved BASTA selection (2 and 3 mg/L) and was subsequently confirmed through PCR analysis. The transformation efficiencies of NR-421 and FSD-2008 were 0.4% and 0.3%, respectively. The qRT-PCR analysis under stress conditions showed a 13.5-fold higher expression of the IPT gene in T2 transgenic plants of NR-421 and a 5.8-fold higher expression in those of FSD-2008 than in non-transgenic controls. Under stress conditions, the wheat transgenic plants exhibited increased chlorophyll and relative water content. Additionally, for total soluble proteins, two transgenic lines from the NR-421 variety showed a significant increase, whereas no notable change was observed in the FSD-2008 transgenics. Moreover, the transgenic lines displayed increased plant height, higher fresh and dry biomass, and increased seed weight compared to the non-transgenic controls. These findings highlight that stress-inducible expression of the IPT gene in wheat leads to enhanced grain yield and subsequently improved drought tolerance.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"7"},"PeriodicalIF":2.7,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142955703","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Resistance of Populus davidiana × P. bolleana overexpressing cinnamoyl-CoA reductase gene to Lymantria dispar larvae. 山杨的抗性研究。过表达肉桂酰辅酶a还原酶基因对异毒蛾幼虫的影响。

IF 2.7 3区生物学

Transgenic Research Pub Date : 2025-01-09 DOI: 10.1007/s11248-024-00426-5

Ye Li, Ruiqiong Zhang, Lili Sun, Chuanwang Cao

引用次数: 0