Biology Open最新文献_第9页

Short- and long-term exposure to high glucose induces unique transcriptional changes in osteoblasts in vitro. 短期和长期暴露于高葡萄糖可诱导体外成骨细胞发生独特的转录变化。

IF 2.4 4区生物学

Biology Open Pub Date : 2024-05-15 Epub Date: 2024-05-14 DOI: 10.1242/bio.060239

Niki Jalava, Milja Arponen, Nicko Widjaja, Terhi J Heino, Kaisa K Ivaska

{"title":"Short- and long-term exposure to high glucose induces unique transcriptional changes in osteoblasts in vitro.","authors":"Niki Jalava, Milja Arponen, Nicko Widjaja, Terhi J Heino, Kaisa K Ivaska","doi":"10.1242/bio.060239","DOIUrl":"10.1242/bio.060239","url":null,"abstract":"Bone is increasingly recognized as a target for diabetic complications. In order to evaluate the direct effects of high glucose on bone, we investigated the global transcriptional changes induced by hyperglycemia in osteoblasts in vitro. Rat bone marrow-derived mesenchymal stromal cells were differentiated into osteoblasts for 10 days, and prior to analysis, they were exposed to hyperglycemia (25 mM) for the short-term (1 or 3 days) or long-term (10 days). Genes and pathways regulated by hyperglycemia were identified using mRNA sequencing and verified with qPCR. Genes upregulated by 1-day hyperglycemia were, for example, related to extracellular matrix organization, collagen synthesis and bone formation. This stimulatory effect was attenuated by 3 days. Long-term exposure impaired osteoblast viability, and downregulated, for example, extracellular matrix organization and lysosomal pathways, and increased intracellular oxidative stress. Interestingly, transcriptional changes by different exposure times were mostly unique and only 89 common genes responding to glucose were identified. In conclusion, short-term hyperglycemia had a stimulatory effect on osteoblasts and bone formation, whereas long-term hyperglycemia had a negative effect on intracellular redox balance, osteoblast viability and function.","PeriodicalId":9216,"journal":{"name":"Biology Open","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11128269/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140916164","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A methodological approach for collecting simultaneous measures of muscle, aponeurosis, and tendon behaviour during dynamic contractions. 在动态收缩过程中同时测量肌肉、肌腱和肌腱行为的方法。

IF 2.4 4区生物学

Biology Open Pub Date : 2024-05-15 Epub Date: 2024-05-23 DOI: 10.1242/bio.060383

Stephanie A Ross, Christine Waters-Banker, Andrew Sawatsky, Timothy R Leonard, Walter Herzog

{"title":"A methodological approach for collecting simultaneous measures of muscle, aponeurosis, and tendon behaviour during dynamic contractions.","authors":"Stephanie A Ross, Christine Waters-Banker, Andrew Sawatsky, Timothy R Leonard, Walter Herzog","doi":"10.1242/bio.060383","DOIUrl":"10.1242/bio.060383","url":null,"abstract":"Skeletal muscles and the tendons that attach them to bone are structurally complex and deform non-uniformly during contraction. While these tissue deformations dictate force production during movement, our understanding of this behaviour is limited due to challenges in obtaining complete measures of the constituent structures. To address these challenges, we present an approach for simultaneously measuring muscle, fascicle, aponeurosis, and tendon behaviour using sonomicrometry. To evaluate this methodology, we conducted isometric and dynamic contractions in in situ rabbit medial gastrocnemius. We found comparable patterns of strain in the muscle belly, fascicle, aponeurosis, and tendon during the isometric trials to those published in the literature. For the dynamic contractions, we found that our measures using this method were consistent across all animals and aligned well with our theoretical understanding of muscle-tendon unit behaviour. Thus, this method provides a means to fully capture the complex behaviour of muscle-tendon units across contraction types.","PeriodicalId":9216,"journal":{"name":"Biology Open","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11139038/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141080041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Climate change consequences on the systemic heart of female Octopus maya: oxidative phosphorylation assessment and the antioxidant system. 气候变化对雌性玛雅章鱼系统性心脏的影响：氧化磷酸化评估和抗氧化系统。

IF 2.4 4区生物学

Biology Open Pub Date : 2024-05-15 Epub Date: 2024-05-16 DOI: 10.1242/bio.060103

Ana Karen Meza-Buendia, Omar Emiliano Aparicio-Trejo, Fernando Díaz, José Pedraza-Chaverri, Carolina Álvarez-Delgado, Carlos Rosas

{"title":"Climate change consequences on the systemic heart of female Octopus maya: oxidative phosphorylation assessment and the antioxidant system.","authors":"Ana Karen Meza-Buendia, Omar Emiliano Aparicio-Trejo, Fernando Díaz, José Pedraza-Chaverri, Carolina Álvarez-Delgado, Carlos Rosas","doi":"10.1242/bio.060103","DOIUrl":"10.1242/bio.060103","url":null,"abstract":"There is evidence that indicates that temperature modulates the reproduction of the tropical species Octopus maya, through the over- or under-expression of many genes in the brain. If the oxygen supply to the brain depends on the circulatory system, how temperature affects different tissues will begin in the heart, responsible for pumping the oxygen to tissues. The present study examines the impact of heat stress on the mitochondrial function of the systemic heart of adult O. maya. The mitochondrial metabolism and antioxidant defense system were measured in the systemic heart tissue of female organisms acclimated to different temperatures (24, 26, and 30°C). The results show that acclimation temperature affects respiratory State 3 and State 4o (oligomycin-induced) with higher values observed in females acclimated at 26°C. The antioxidant defense system is also affected by acclimation temperature with significant differences observed in superoxide dismutase, glutathione S-transferase activities, and glutathione levels. The results suggest that high temperatures (30°C) could exert physical limitations on the circulatory system through the heart pumping, affecting nutrient and oxygen transport to other tissues, including the brain, which exerts control over the reproductive system. The role of the cardiovascular system in supporting aerobic metabolism in octopus females is discussed.","PeriodicalId":9216,"journal":{"name":"Biology Open","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11155352/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140944057","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Na,K-ATPase activity promotes macropinocytosis in colon cancer via Wnt signaling. Na,K-ATP酶活性通过Wnt信号传导促进结肠癌中的大蛋白细胞增殖

IF 1.8 4区生物学

Biology Open Pub Date : 2024-05-15 Epub Date: 2024-05-21 DOI: 10.1242/bio.060269

Nydia Tejeda-Muñoz, Yagmur Azbazdar, Eric A Sosa, Julia Monka, Pu-Sheng Wei, Grace Binder, Kuo-Ching Mei, Yerbol Z Kurmangaliyev, Edward M De Robertis

{"title":"Na,K-ATPase activity promotes macropinocytosis in colon cancer via Wnt signaling.","authors":"Nydia Tejeda-Muñoz, Yagmur Azbazdar, Eric A Sosa, Julia Monka, Pu-Sheng Wei, Grace Binder, Kuo-Ching Mei, Yerbol Z Kurmangaliyev, Edward M De Robertis","doi":"10.1242/bio.060269","DOIUrl":"10.1242/bio.060269","url":null,"abstract":"Recent research has shown that membrane trafficking plays an important role in canonical Wnt signaling through sequestration of the β-catenin destruction complex inside multivesicular bodies (MVBs) and lysosomes. In this study, we introduce Ouabain, an inhibitor of the Na,K-ATPase pump that establishes electric potentials across membranes, as a potent inhibitor of Wnt signaling. We find that Na,K-ATPase levels are elevated in advanced colon carcinoma, that this enzyme is elevated in cancer cells with constitutively activated Wnt pathway and is activated by GSK3 inhibitors that increase macropinocytosis. Ouabain blocks macropinocytosis, which is an essential step in Wnt signaling, probably explaining the strong effects of Ouabain on this pathway. In Xenopus embryos, brief Ouabain treatment at the 32-cell stage, critical for the earliest Wnt signal in development-inhibited brains, could be reversed by treatment with Lithium chloride, a Wnt mimic. Inhibiting membrane trafficking may provide a way of targeting Wnt-driven cancers.","PeriodicalId":9216,"journal":{"name":"Biology Open","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2024-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11139033/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140856216","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Short- and long-term exposure to high glucose induces unique transcriptional changes in osteoblasts in vitro. 短期和长期暴露于高葡萄糖可诱导体外成骨细胞发生独特的转录变化。

IF 2.4 4区生物学

Biology Open Pub Date : 2024-05-15 Epub Date: 2024-05-14 DOI: 10.1242/bio.060239

Niki Jalava, Milja Arponen, Nicko Widjaja, Terhi J Heino, Kaisa K Ivaska

{"title":"Short- and long-term exposure to high glucose induces unique transcriptional changes in osteoblasts in vitro.","authors":"Niki Jalava, Milja Arponen, Nicko Widjaja, Terhi J Heino, Kaisa K Ivaska","doi":"10.1242/bio.060239","DOIUrl":"10.1242/bio.060239","url":null,"abstract":"Bone is increasingly recognized as a target for diabetic complications. In order to evaluate the direct effects of high glucose on bone, we investigated the global transcriptional changes induced by hyperglycemia in osteoblasts in vitro. Rat bone marrow-derived mesenchymal stromal cells were differentiated into osteoblasts for 10 days, and prior to analysis, they were exposed to hyperglycemia (25 mM) for the short-term (1 or 3 days) or long-term (10 days). Genes and pathways regulated by hyperglycemia were identified using mRNA sequencing and verified with qPCR. Genes upregulated by 1-day hyperglycemia were, for example, related to extracellular matrix organization, collagen synthesis and bone formation. This stimulatory effect was attenuated by 3 days. Long-term exposure impaired osteoblast viability, and downregulated, for example, extracellular matrix organization and lysosomal pathways, and increased intracellular oxidative stress. Interestingly, transcriptional changes by different exposure times were mostly unique and only 89 common genes responding to glucose were identified. In conclusion, short-term hyperglycemia had a stimulatory effect on osteoblasts and bone formation, whereas long-term hyperglycemia had a negative effect on intracellular redox balance, osteoblast viability and function.","PeriodicalId":9216,"journal":{"name":"Biology Open","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141160457","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Arf GTPase-Activating proteins ADAP1 and ARAP1 regulate incorporation of CD63 in multivesicular bodies. Arf GTP酶激活蛋白ADAP1和ARAP1调节CD63在多囊体中的结合。

IF 1.8 4区生物学

Biology Open Pub Date : 2024-05-15 Epub Date: 2024-05-10 DOI: 10.1242/bio.060338

Kasumi Suzuki, Yoshitaka Okawa, Sharmin Akter, Haruki Ito, Yoko Shiba

{"title":"Arf GTPase-Activating proteins ADAP1 and ARAP1 regulate incorporation of CD63 in multivesicular bodies.","authors":"Kasumi Suzuki, Yoshitaka Okawa, Sharmin Akter, Haruki Ito, Yoko Shiba","doi":"10.1242/bio.060338","DOIUrl":"10.1242/bio.060338","url":null,"abstract":"Arf GTPase-activating proteins (ArfGAPs) mediate the hydrolysis of GTP bound to ADP-ribosylation factors. ArfGAPs are critical for cargo sorting in the Golgi-to-ER traffic. However, the role of ArfGAPs in sorting into intralumenal vesicles (ILVs) in multivesicular bodies (MVBs) in post-Golgi traffic remains unclear. Exosomes are extracellular vesicles (EVs) of endosomal origin. CD63 is an EV marker. CD63 is enriched ILVs in MVBs of cells. However, the secretion of CD63 positive EVs has not been consistent with the data on CD63 localization in MVBs, and how CD63-containing EVs are formed is yet to be understood. To elucidate the mechanism of CD63 transport to ILVs, we focused on CD63 localization in MVBs and searched for the ArfGAPs involved in CD63 localization. We observed that ADAP1 and ARAP1 depletion inhibited CD63 localization to enlarged endosomes after Rab5Q79L overexpression. We tested epidermal growth factor (EGF) and CD9 localization in MVBs. We observed that ADAP1 and ARAP1 depletion inhibited CD9 localization in enlarged endosomes but not EGF. Our results indicate ADAP1 and ARAP1, regulate incorporation of CD63 and CD9, but not EGF, in overlapped and different MVBs. Our work will contribute to distinguish heterogenous ILVs and exosomes by ArfGAPs.","PeriodicalId":9216,"journal":{"name":"Biology Open","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2024-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11103404/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140859738","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Vascular smooth muscle cell-derived nerve growth factor regulates sympathetic collateral branching to innervate blood vessels in embryonic skin. 血管平滑肌细胞源性神经生长因子调节交感神经侧枝，以支配胚胎皮肤中的血管。

IF 2.4 4区生物学

Biology Open Pub Date : 2024-04-19 DOI: 10.1242/bio.060147

Wenling Li, Katherine Lipsius, Nathan G Burns, Ryo Sato, Azaan Rehman, Hui Xue, Christian Combs, Liliana Minichiello, Harshi Gangrade, Emmanouil Tampakakis, Y. Mukouyama

{"title":"Vascular smooth muscle cell-derived nerve growth factor regulates sympathetic collateral branching to innervate blood vessels in embryonic skin.","authors":"Wenling Li, Katherine Lipsius, Nathan G Burns, Ryo Sato, Azaan Rehman, Hui Xue, Christian Combs, Liliana Minichiello, Harshi Gangrade, Emmanouil Tampakakis, Y. Mukouyama","doi":"10.1242/bio.060147","DOIUrl":"https://doi.org/10.1242/bio.060147","url":null,"abstract":"Blood vessels serve as intermediate conduits for the extension of sympathetic axons towards target tissues, while also acting as crucial targets for their homeostatic processes encompassing the regulation of temperature, blood pressure, and oxygen availability. How sympathetic axons innervate not only blood vessels but also a wide array of target tissues is not clear. Here we show that in embryonic skin, after the establishment of co-branching between sensory nerves and blood vessels, sympathetic axons invade the skin alongside these sensory nerves and extend their branches towards these blood vessels covered by vascular smooth muscle cells (VSMCs). Our mosaic labeling technique for sympathetic axons shows that collateral branching predominantly mediates the innervation of VSMC-covered blood vessels by sympathetic axons. The expression of nerve growth factor (NGF), previously known to induce collateral axon branching in culture, can be detected in the vascular smooth muscle cell (VSMC)-covered blood vessels, as well as sensory nerves. Indeed, VSMC-specific Ngf knockout leads to a significant decrease of collateral branching of sympathetic axons innervating VSMC-covered blood vessels. These data suggest that VSMC-derived NGF serves as an inductive signal for collateral branching of sympathetic axons innervating blood vessels in the embryonic skin.","PeriodicalId":9216,"journal":{"name":"Biology Open","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140683779","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Biomechanical analysis of little penguins' underwater locomotion from the free-ranging dive data. 根据自由潜水数据对小企鹅水下运动进行生物力学分析。

IF 2.4 4区生物学

Biology Open Pub Date : 2024-04-19 DOI: 10.1242/bio.060244

M. Masud, Peter Dabnichki

{"title":"Biomechanical analysis of little penguins' underwater locomotion from the free-ranging dive data.","authors":"M. Masud, Peter Dabnichki","doi":"10.1242/bio.060244","DOIUrl":"https://doi.org/10.1242/bio.060244","url":null,"abstract":"Penguins are proficient swimmers, and their survival depends on their ability to catch prey. The diving behaviour of these fascinating birds should then minimize the associated energy cost. For the first time, the energy cost of penguin dives is computed from the free-ranging dive data, on the basis of an existing biomechanical model. Time-resolved acceleration and depth data collected for 300 dives of little penguins (Eudyptula minor) are specifically employed to compute the bird dive angles and swimming speeds, which are needed for the energy estimate. We find that the numerically obtained energy cost by using the free-ranging dive data is not far from the minimum cost predicted by the model. The outcome, therefore, supports the physical soundness of the chosen model; however, it also suggests that, for closer agreement, one should consider previously neglected effects, such as those due to water currents and those associated with motion unsteadiness. Additionally, from the free-ranging dive data, we calculate hydrodynamic forces and non-dimensional indicators of propulsion performance - Strouhal and Reynolds numbers. The obtained values further confirm that little penguins employ efficient propulsion mechanisms, in agreement with previous investigations.","PeriodicalId":9216,"journal":{"name":"Biology Open","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140685461","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Cdc48 and its co-factor Ufd1 extract CENP-A from centromeric chromatin and can induce chromosome elimination in the fission yeast Schizosaccharomyces pombe. Cdc48 及其辅助因子 Ufd1 能从中心染色质中提取 CENP-A，并能诱导裂殖酵母中染色体的消亡。

IF 2.4 4区生物学

Biology Open Pub Date : 2024-04-15 Epub Date: 2024-04-08 DOI: 10.1242/bio.060287

Yukiko Nakase, Hiroaki Murakami, Michiko Suma, Kaho Nagano, Airi Wakuda, Teppei Kitagawa, Tomohiro Matsumoto

{"title":"Cdc48 and its co-factor Ufd1 extract CENP-A from centromeric chromatin and can induce chromosome elimination in the fission yeast Schizosaccharomyces pombe.","authors":"Yukiko Nakase, Hiroaki Murakami, Michiko Suma, Kaho Nagano, Airi Wakuda, Teppei Kitagawa, Tomohiro Matsumoto","doi":"10.1242/bio.060287","DOIUrl":"10.1242/bio.060287","url":null,"abstract":"CENP-A determines the identity of the centromere. Because the position and size of the centromere and its number per chromosome must be maintained, the distribution of CENP-A is strictly regulated. In this study, we have aimed to understand mechanisms to regulate the distribution of CENP-A (Cnp1SP) in fission yeast. A mutant of the ufd1+ gene (ufd1-73) encoding a cofactor of Cdc48 ATPase is sensitive to Cnp1 expressed at a high level and allows mislocalization of Cnp1. The level of Cnp1 in centromeric chromatin is increased in the ufd1-73 mutant even when Cnp1 is expressed at a normal level. A preexisting mutant of the cdc48+ gene (cdc48-353) phenocopies the ufd1-73 mutant. We have also shown that Cdc48 and Ufd1 proteins interact physically with centromeric chromatin. Finally, Cdc48 ATPase with Ufd1 artificially recruited to the centromere of a mini-chromosome (Ch16) induce a loss of Cnp1 from Ch16, leading to an increased rate of chromosome loss. It appears that Cdc48 ATPase, together with its cofactor Ufd1 remove excess Cnp1 from chromatin, likely in a direct manner. This mechanism may play a role in centromere disassembly, a process to eliminate Cnp1 to inactivate the kinetochore function during development, differentiation, and stress response.","PeriodicalId":9216,"journal":{"name":"Biology Open","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11033524/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140206369","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Efficient genome editing using modified Cas9 proteins in zebrafish. 利用改良 Cas9 蛋白对斑马鱼进行高效基因组编辑。

IF 2.4 4区生物学

Biology Open Pub Date : 2024-04-15 Epub Date: 2024-03-28 DOI: 10.1242/bio.060401

Laura Dorner, Benedikt Stratmann, Laura Bader, Marco Podobnik, Uwe Irion

{"title":"Efficient genome editing using modified Cas9 proteins in zebrafish.","authors":"Laura Dorner, Benedikt Stratmann, Laura Bader, Marco Podobnik, Uwe Irion","doi":"10.1242/bio.060401","DOIUrl":"10.1242/bio.060401","url":null,"abstract":"The zebrafish (Danio rerio) is an important model organism for basic as well as applied bio-medical research. One main advantage is its genetic tractability, which was greatly enhanced by the introduction of the CRISPR/Cas method a decade ago. The generation of loss-of-function alleles via the production of small insertions or deletions in the coding sequences of genes with CRISPR/Cas systems is now routinely achieved with high efficiency. The method is based on the error prone repair of precisely targeted DNA double strand breaks by non-homologous end joining (NHEJ) in the cell nucleus. However, editing the genome with base pair precision, by homology-directed repair (HDR), is by far less efficient and therefore often requires large-scale screening of potential carriers by labour intensive genotyping. Here we confirm that the Cas9 protein variant SpRY, with relaxed PAM requirement, can be used to target some sites in the zebrafish genome. In addition, we demonstrate that the incorporation of an artificial nuclear localisation signal (aNLS) into the Cas9 protein variants not only enhances the efficiency of gene knockout but also the frequency of HDR, thereby facilitating the efficient modification of single base pairs in the genome. Our protocols provide a guide for a cost-effective generation of versatile and potent Cas9 protein variants and efficient gene editing in zebrafish.","PeriodicalId":9216,"journal":{"name":"Biology Open","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10997048/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140304948","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0