Journal of Biological Engineering最新文献

{"title":"Recent advances in 3D printing of biodegradable metals for orthopaedic applications.","authors":"Wenqing Liang, Chao Zhou, Hongwei Zhang, Juqin Bai, Bo Jiang, Chanyi Jiang, Wenyi Ming, Hengjian Zhang, Hengguo Long, Xiaogang Huang, Jiayi Zhao","doi":"10.1186/s13036-023-00371-7","DOIUrl":"10.1186/s13036-023-00371-7","url":null,"abstract":"<p><p>The use of biodegradable polymers for treating bone-related diseases has become a focal point in the field of biomedicine. Recent advancements in material technology have expanded the range of materials suitable for orthopaedic implants. Three-dimensional (3D) printing technology has become prevalent in healthcare, and while organ printing is still in its early stages and faces ethical and technical hurdles, 3D printing is capable of creating 3D structures that are supportive and controllable. The technique has shown promise in fields such as tissue engineering and regenerative medicine, and new innovations in cell and bio-printing and printing materials have expanded its possibilities. In clinical settings, 3D printing of biodegradable metals is mainly used in orthopedics and stomatology. 3D-printed patient-specific osteotomy instruments, orthopedic implants, and dental implants have been approved by the US FDA for clinical use. Metals are often used to provide support for hard tissue and prevent complications. Currently, 70-80% of clinically used implants are made from niobium, tantalum, nitinol, titanium alloys, cobalt-chromium alloys, and stainless steels. However, there has been increasing interest in biodegradable metals such as magnesium, calcium, zinc, and iron, with numerous recent findings. The advantages of 3D printing, such as low manufacturing costs, complex geometry capabilities, and short fabrication periods, have led to widespread adoption in academia and industry. 3D printing of metals with controllable structures represents a cutting-edge technology for developing metallic implants for biomedical applications. This review explores existing biomaterials used in 3D printing-based orthopedics as well as biodegradable metals and their applications in developing metallic medical implants and devices. The challenges and future directions of this technology are also discussed.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"56"},"PeriodicalIF":5.6,"publicationDate":"2023-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10466721/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10500355","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimizing the fabrication of a 3D high-resolution implant for neural stimulation.","authors":"Gal Shpun, Nairouz Farah, Yoav Chemla, Amos Markus, Tamar Azrad Leibovitch, Erel Lasnoy, Doron Gerber, Zeev Zalevsky, Yossi Mandel","doi":"10.1186/s13036-023-00370-8","DOIUrl":"10.1186/s13036-023-00370-8","url":null,"abstract":"<p><strong>Background: </strong>Tissue-integrated micro-electronic devices for neural stimulation hold great potential in restoring the functionality of degenerated organs, specifically, retinal prostheses, which are aimed at vision restoration. The fabrication process of 3D polymer-metal devices with high resolution and a high aspect-ratio (AR) is very complex and faces many challenges that impair its functionality.</p><p><strong>Approach: </strong>Here we describe the optimization of the fabrication process of a bio-functionalized 3D high-resolution 1mm circular subretinal implant composed of SU-8 polymer integrated with dense gold microelectrodes (23μm pitch) passivated with 3D micro-well-like structures (20μm diameter, 3μm resolution). The main challenges were overcome by step-by-step planning and optimization while utilizing a two-step bi-layer lift-off process; bio-functionalization was carried out by N₂ plasma treatment and the addition of a bio-adhesion molecule.</p><p><strong>Main results: </strong>In-vitro and in-vivo investigations, including SEM and FIB cross section examinations, revealed a good structural design, as well as a good long-term integration of the device in the rat sub-retinal space and cell migration into the wells. Moreover, the feasibility of subretinal neural stimulation using the fabricated device was demonstrated in-vitro by electrical activation of rat's retina.</p><p><strong>Conclusions: </strong>The reported process and optimization steps described here in detail can aid in designing and fabricating retinal prosthetic devices or similar neural implants.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"55"},"PeriodicalIF":5.6,"publicationDate":"2023-08-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10463680/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10174274","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel inductively coupled capacitor wireless sensor system for rapid antibiotic susceptibility testing.","authors":"Yikang Xu, Dacheng Ren","doi":"10.1186/s13036-023-00373-5","DOIUrl":"10.1186/s13036-023-00373-5","url":null,"abstract":"<p><strong>Background: </strong>The increasing prevalence and severity of antimicrobial resistance (AMR) present a major challenge to our healthcare system. Rapid detection of AMR is essential for lifesaving under emergent conditions such as sepsis. The current gold standard phenotypic antibiotic susceptibility testing (AST) takes more than a day to obtain results. Genotypic ASTs are faster (hours) in detecting the presence of resistance genes but require specific probes/knowledge of each AMR gene and do not provide specific information at the phenotype level. To address this unmet challenge, we developed a new rapid phenotypic AST.</p><p><strong>Result: </strong>We designed a new electrochemical biosensor based on the concept of magnetically coupled LC sensors. The engineered LC sensors can be placed in 96-well plates and communicate the reading remotely with a receiver coil for signal analysis. The sensors were validated by monitoring the growth of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa in the presence and absence of different antibiotics. Drug-resistant strains were used as controls. Bacterial growth was detected within 30 min after inoculation, allowing rapid determination of antibiotic susceptibility at the phenotype level. The sensor also functions in the presence of host proteins when tested with 2% FBS in growth media.</p><p><strong>Conclusions: </strong>With the compatibility with 96-well plates, this label-free rapid 30-min AST has the potential for low-cost applications with simple integration into the existing workflow in clinical settings.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"54"},"PeriodicalIF":5.6,"publicationDate":"2023-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10439655/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10048338","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cancer-on-chip: a 3D model for the study of the tumor microenvironment.","authors":"Elisa Cauli, Michela Anna Polidoro, Simona Marzorati, Claudio Bernardi, Marco Rasponi, Ana Lleo","doi":"10.1186/s13036-023-00372-6","DOIUrl":"10.1186/s13036-023-00372-6","url":null,"abstract":"<p><p>The approval of anticancer therapeutic strategies is still slowed down by the lack of models able to faithfully reproduce in vivo cancer physiology. On one hand, the conventional in vitro models fail to recapitulate the organ and tissue structures, the fluid flows, and the mechanical stimuli characterizing the human body compartments. On the other hand, in vivo animal models cannot reproduce the typical human tumor microenvironment, essential to study cancer behavior and progression. This study reviews the cancer-on-chips as one of the most promising tools to model and investigate the tumor microenvironment and metastasis. We also described how cancer-on-chip devices have been developed and implemented to study the most common primary cancers and their metastatic sites. Pros and cons of this technology are then discussed highlighting the future challenges to close the gap between the pre-clinical and clinical studies and accelerate the approval of new anticancer therapies in humans.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"53"},"PeriodicalIF":5.6,"publicationDate":"2023-08-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10436436/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10047326","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fluid dynamic design for mitigating undesired cell effects and its application to testis cell response testing to endocrine disruptors.","authors":"Seungjin Lee, Jinseop Ahn, Seok-Man Kim, Daehan Kim, Jiun Yeom, Jeongmok Kim, Joong Yull Park, Buom-Yong Ryu","doi":"10.1186/s13036-023-00369-1","DOIUrl":"10.1186/s13036-023-00369-1","url":null,"abstract":"<p><p>Microfluidic devices have emerged as powerful tools for cell-based experiments, offering a controlled microenvironment that mimic the conditions within the body. Numerous cell experiment studies have successfully utilized microfluidic channels to achieve various new scientific discoveries. However, it has been often overlooked that undesired and unnoticed propagation of cellular molecules in such bio-microfluidic channel systems can have a negative impact on the experimental results. Thus, more careful designing is required to minimize such unwanted issues through deeper understanding and careful control of chemically and physically predominant factors at the microscopic scale. In this paper, we introduce a new approach to improve microfluidic channel design, specifically targeting the mitigation of the aforementioned challenges. To minimize the occurrence of undesired cell positioning upstream from the main test section where a concentration gradient field locates, an additional narrow port structure was devised between the microfluidic upstream channel and each inlet reservoir. This port also functioned as a passive lock that hold the flow at rest via fluid-air surface tension, which facilitated manual movement of the device even when cell attachment was not achieved completely. To demonstrate the practicability of the system, we conducted experiments and diffusion simulations on the effect of endocrine disruptors on germ cells. To this end, a bisphenol-A (BPA) concentration gradient was generated in the main channel of the system at BPA concentrations ranging from 120.8 μM to 79.3 μM, and the proliferation of GC-1 cells in the BPA gradient environment was quantitatively evaluated. The features and concepts of the introduced design is to minimize unexpected and ignored error sources, which will be one of the issues to be considered in the development of microfluidic systems to explore extremely delicate cellular phenomena.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"51"},"PeriodicalIF":5.6,"publicationDate":"2023-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10408176/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10319206","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tissue poromechanical deformation effects on steam pop likelihood in 3-D radiofrequency cardiac ablation.","authors":"Patcharaporn Wongchadakul, Ashim K Datta, Phadungsak Rattanadecho","doi":"10.1186/s13036-023-00365-5","DOIUrl":"10.1186/s13036-023-00365-5","url":null,"abstract":"<p><p>Radiofrequency Cardiac Ablation (RFCA) is a common procedure that heats cardiac tissue to destroy abnormal signal pathways to eliminate arrhythmias. The complex multiphysics phenomena during this procedure need to be better understood to improve both procedure and device design. A deformable poromechanical model of cardiac tissue was developed that coupled joule heating from the electrode, heat transfer, and blood flow from normal perfusion and thermally driven natural convection, which mimics the real tissue structure more closely and provides more realistic results compared to previous models. The expansion of tissue from temperature rise reduces blood velocity, leading to increased tissue temperature, thus affecting steam pop occurrence. Detailed temperature velocity, and thermal expansion of the tissue provided a comprehensive picture of the process. Poromechanical expansion of the tissue from temperature rise reduces blood velocity, increasing tissue temperature. Tissue properties influence temperatures, with lower porosity increasing the temperatures slightly, due to lower velocities. Deeper electrode insertion raises temperature due to increased current flow. The results demonstrate that a 5% increase in porosity leads to a considerable 10% increase in maximum tissue temperature. These insights should greatly help in avoiding undesirable heating effects that can lead to steam pop and in designing improved electrodes.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"52"},"PeriodicalIF":5.6,"publicationDate":"2023-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10408080/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9956190","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative transcriptome findings reveal the neuroinflammatory network and potential biomarkers to early detection of ischemic stroke.","authors":"Jiefeng Luo,&nbsp;Dingzhi Chen,&nbsp;Yujia Mei,&nbsp;Hepeng Li,&nbsp;Biyun Qin,&nbsp;Xiao Lin,&nbsp;Ting Fung Chan,&nbsp;Keng Po Lai,&nbsp;Deyan Kong","doi":"10.1186/s13036-023-00362-8","DOIUrl":"https://doi.org/10.1186/s13036-023-00362-8","url":null,"abstract":"<p><strong>Introduction: </strong>Ischemic stroke accounts for 70-80% of all stroke cases, leading to over two million people dying every year. Poor diagnosis and late detection are the major causes of the high death and disability rate.</p><p><strong>Methods: </strong>In the present study, we used the middle cerebral artery occlusion (MCAO) rat model and applied comparative transcriptomic analysis, followed by a systematic advanced bioinformatic analysis, including gene ontology enrichment analysis and Ingenuity Pathway Analysis (IPA). We aimed to identify novel biomarkers for the early detection of ischemic stroke. In addition, we aimed to delineate the molecular mechanisms underlying the development of ischemic stroke, in which we hoped to identify novel therapeutic targets for treating ischemic stroke.</p><p><strong>Results: </strong>In the comparative transcriptomic analysis, we identified 2657 differentially expressed genes (DEGs) in the brain tissue of the MCAO model. The gene enrichment analysis highlighted the importance of these DEGs in oxygen regulation, neural functions, and inflammatory and immune responses. We identified the elevation of angiopoietin-2 and leptin receptor as potential novel biomarkers for early detection of ischemic stroke. Furthermore, the result of IPA suggested targeting the inflammasome pathway, integrin-linked kinase signaling pathway, and Th1 signaling pathway for treating ischemic stroke.</p><p><strong>Conclusion: </strong>The results of the present study provide novel insight into the biomarkers and therapeutic targets as potential treatments of ischemic stroke.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"50"},"PeriodicalIF":5.6,"publicationDate":"2023-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10398984/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9935281","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Non-viral gene delivery to human mesenchymal stem cells: a practical guide towards cell engineering.","authors":"Natalia Carballo-Pedrares, Federica Ponti, Junquera Lopez-Seijas, Diego Miranda-Balbuena, Nina Bono, Gabriele Candiani, Ana Rey-Rico","doi":"10.1186/s13036-023-00363-7","DOIUrl":"10.1186/s13036-023-00363-7","url":null,"abstract":"<p><p>In recent decades, human mesenchymal stem cells (hMSCs) have gained momentum in the field of cell therapy for treating cartilage and bone injuries. Despite the tri-lineage multipotency, proliferative properties, and potent immunomodulatory effects of hMSCs, their clinical potential is hindered by donor variations, limiting their use in medical settings. To address this challenge, gene delivery technologies have emerged as a promising approach to modulate the phenotype and commitment of hMSCs towards specific cell lineages, thereby enhancing osteochondral repair strategies. This review provides a comprehensive overview of current non-viral gene delivery approaches used to engineer MSCs, highlighting key factors such as the choice of nucleic acid or delivery vector, transfection strategies, and experimental parameters. Additionally, it outlines various protocols and methods for qualitative and quantitative evaluation of their therapeutic potential as a delivery system in osteochondral regenerative applications. In summary, this technical review offers a practical guide for optimizing non-viral systems in osteochondral regenerative approaches. hMSCs constitute a key target population for gene therapy techniques. Nevertheless, there is a long way to go for their translation into clinical treatments. In this review, we remind the most relevant transfection conditions to be optimized, such as the type of nucleic acid or delivery vector, the transfection strategy, and the experimental parameters to accurately evaluate a delivery system. This survey provides a practical guide to optimizing non-viral systems for osteochondral regenerative approaches.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"49"},"PeriodicalIF":5.7,"publicationDate":"2023-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10369726/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9936040","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An injectable, self-healing, electroconductive hydrogel loaded with neural stem cells and donepezil for enhancing local therapy effect of spinal cord injury.","authors":"Tiemei Liu,&nbsp;Qiang Zhang,&nbsp;Hongru Li,&nbsp;Xiaoqian Cui,&nbsp;Zhiping Qi,&nbsp;Xiaoyu Yang","doi":"10.1186/s13036-023-00368-2","DOIUrl":"https://doi.org/10.1186/s13036-023-00368-2","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Spinal cord injury (SCI) is a serious injury with high mortality and disability rates, and there is no effective treatment at present. It has been reported that some treatments, such as drug intervention and stem cell transplantation have positive effects in promoting neurological recovery. Although those treatments are effective for nerve regeneration, many drawbacks, such as low stem cell survival rates and side effects caused by systemic medication, have limited their development. In recent years, injectable hydrogel materials have been widely used in tissue engineering due to their good biocompatibility, biodegradability, controllable properties, and low invasiveness. The treatment strategy of injectable hydrogels combined with stem cells or drugs has made some progress in SCI repair, showing the potential to overcome the drawbacks of traditional drugs and stem cell therapy.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;In this study, a novel injectable electroactive hydrogel (NGP) based on sodium hyaluronate oxide (SAO) and polyaniline-grafted gelatine (NH&lt;sub&gt;2&lt;/sub&gt;-Gel-PANI) was developed as a material in which to load neural stem cells (NSCs) and donepezil (DPL) to facilitate nerve regeneration after SCI. To evaluate the potential of the prepared NGP hydrogel in SCI repair applications, the surface morphology, self-repairing properties, electrical conductivity and cytocompatibility of the resulting hydrogel were analysed. Meanwhile, we evaluated the neural repair ability of NGP hydrogels loaded with DPL and NSCs using a rat model of spinal cord injury.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;The NGP hydrogel has a suitable pore size, good biocompatibility, excellent conductivity, and injectable and self-repairing properties, and its degradation rate matches the repair cycle of spinal cord injury. In addition, DPL could be released continuously and slowly from the NGP hydrogel; thus, the NGP hydrogel could serve as an excellent carrier for drugs and cells. The results of in vitro cell experiments showed that the NGP hydrogel had good cytocompatibility and could significantly promote the neuronal differentiation and axon growth of NSCs, and loading the hydrogel with DPL could significantly enhance this effect. More importantly, the NGP hydrogel loaded with DPL showed a significant inhibitory effect on astrocytic differentiation of NSCs in vitro. Animal experiments showed that the combination of NGP hydrogel, DPL, and NSCs had the best therapeutic effect on the recovery of motor function and nerve conduction function in rats. NGP hydrogel loaded with NSCs and DPL not only significantly increased the myelin sheath area, number of new neurons and axon area but also minimized the area of the cystic cavity and glial scar and promoted neural circuit reconstruction.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusions: &lt;/strong&gt;The DPL- and NSC-laden electroactive hydrogel developed in this study is an ideal biomaterial for the treatment of traumatic spinal cord ","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"48"},"PeriodicalIF":5.6,"publicationDate":"2023-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10367392/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10232952","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Study of the regulatory elements of the Ovalbumin gene promoter using CRISPR technology in chicken cells.","authors":"Sara Yousefi Taemeh,&nbsp;Nima Dehdilani,&nbsp;Lena Goshayeshi,&nbsp;Sylvie Rival-Gervier,&nbsp;Jalil Mehrzad,&nbsp;Bertrand Pain,&nbsp;Hesam Dehghani","doi":"10.1186/s13036-023-00367-3","DOIUrl":"https://doi.org/10.1186/s13036-023-00367-3","url":null,"abstract":"<p><strong>Background: </strong>Hormone-dependent promoters are very efficient in transgene expression. Plasmid-based reporter assays have identified regulatory sequences of the Ovalbumin promoter that are involved in response to estrogen and have shown that the deletion of the steroid-dependent regulatory element (SDRE) and negative regulatory element (NRE) leads to a steroid-independent expression of a reporter. However, the functional roles of these regulatory elements within the native genomic context of the Ovalbumin promoter have not been evaluated.</p><p><strong>Results: </strong>In this study, we show that the negative effects of the NRE element on the Ovalbumin gene can be counteracted by CRISPR interference. We also show that the CRISPR-mediated deletion of SDRE and NRE promoter elements in a non-oviduct cell can lead to the significant expression of the Ovalbumin gene. In addition, the targeted knock-in of a transgene reporter in the Ovalbumin coding region and its expression confirms that the truncated promoter of the Ovalbumin gene can be efficiently used for an estrogen-independent expression of a foreign gene.</p><p><strong>Conclusions: </strong>The methodology applied in this paper allowed the study of promoter regulatory sequences in their native nuclear organization.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"46"},"PeriodicalIF":5.6,"publicationDate":"2023-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10353141/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9836289","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}