aBIOTECH最新文献

{"title":"Developing a CRISPR/FrCas9 system for core promoter editing in rice","authors":"Hui Wang,&nbsp;Jian Ding,&nbsp;Jingyan Zhu,&nbsp;Xiaoshuang Liu,&nbsp;Rongfang Xu,&nbsp;Ruiying Qin,&nbsp;Dongfang Gu,&nbsp;Min Li,&nbsp;Pengcheng Wei,&nbsp;Juan Li","doi":"10.1007/s42994-024-00157-5","DOIUrl":"10.1007/s42994-024-00157-5","url":null,"abstract":"<div><p>Small mutations in the core promoter region of a gene may result in substantial changes in expression strengths. However, targeting TA-rich sequences of core promoters may pose a challenge for Cas9 variants such as SpCas9 and other G-rich PAM-compatible Cas9s. In this study, we engineered a unique FrCas9 system derived from <i>Faecalibaculum rodentium</i> for plant genome editing. Our findings indicate that this system is efficient in rice when the TATA sequence is used as a PAM. In addition, FrCas9 demonstrated activity against all 16 possible NNTA PAMs, achieving an efficiency of up to 35.3% in calli and generating homozygous or biallelic mutations in 31.3% of the T₀ transgenic plants. A proof-of-concept experiment to examine editing of the rice <i>WX</i> core promoter confirmed that FrCas9-induced mutations could modify gene expression and amylose content. Multiplex mutations and deletions were produced by bidirectional editing, mediated by FrCas9, using a single palindromic TATA sequence as a PAM. Moreover, we developed FrCas9-derived base editors capable of programmable conversion between A·T and G·C pairs in plants. This study highlights a versatile FrCas9 toolset for plant core promoter editing, offering great potential for the fine-tuning of gene expression and creating of new germplasms.</p></div>","PeriodicalId":53135,"journal":{"name":"aBIOTECH","volume":null,"pages":null},"PeriodicalIF":4.6,"publicationDate":"2024-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s42994-024-00157-5.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140676941","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Natural GmACO1 allelic variations confer drought tolerance and influence nodule formation in soybean","authors":"Zhifang Zhang,&nbsp;Junkui Ma,&nbsp;Xia Yang,&nbsp;Shan Liang,&nbsp;Yucheng Liu,&nbsp;Yaqin Yuan,&nbsp;Qianjin Liang,&nbsp;Yanting Shen,&nbsp;Guoan Zhou,&nbsp;Min Zhang,&nbsp;Zhixi Tian,&nbsp;Shulin Liu","doi":"10.1007/s42994-024-00160-w","DOIUrl":"10.1007/s42994-024-00160-w","url":null,"abstract":"<div><p>Soybean [<i>Glycine max</i> (L.) Merr.] is one of the most important, but a drought-sensitive, crops. Identifying the genes controlling drought tolerance is important in soybean breeding. Here, through a genome-wide association study, we identified one significant association locus, located on chromosome 8, which conferred drought tolerance variations in a natural soybean population. Allelic analysis and genetic validation demonstrated that <i>GmACO1</i>, encoding for a 1-aminocyclopropane-1-carboxylate oxidase, was the causal gene in this association locus, and positively regulated drought tolerance in soybean. Meanwhile, we determined that <i>GmACO1</i> expression was reduced after rhizobial infection, and that <i>GmACO1</i> negatively regulated soybean nodule formation. Overall, our findings provide insights into soybean cultivars for future breeding.</p></div>","PeriodicalId":53135,"journal":{"name":"aBIOTECH","volume":null,"pages":null},"PeriodicalIF":4.6,"publicationDate":"2024-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s42994-024-00160-w.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140688805","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimized protoplast isolation and transfection with a breakpoint: accelerating Cas9/sgRNA cleavage efficiency validation in monocot and dicot","authors":"Debasmita Panda,&nbsp;Subhasis Karmakar,&nbsp;Manaswini Dash,&nbsp;Swagat Kumar Tripathy,&nbsp;Priya Das,&nbsp;Sagar Banerjee,&nbsp;Yiping Qi,&nbsp;Sanghamitra Samantaray,&nbsp;Pradipta Kumar Mohapatra,&nbsp;Mirza J. Baig,&nbsp;Kutubuddin A. Molla","doi":"10.1007/s42994-024-00139-7","DOIUrl":"10.1007/s42994-024-00139-7","url":null,"abstract":"<div><p>The CRISPR-Cas genome editing tools are revolutionizing agriculture and basic biology with their simplicity and precision ability to modify target genomic loci. Software-predicted guide RNAs (gRNAs) often fail to induce efficient cleavage at target loci. Many target loci are inaccessible due to complex chromatin structure. Currently, there is no suitable tool available to predict the architecture of genomic target sites and their accessibility. Hence, significant time and resources are spent on performing editing experiments with inefficient guides. Although in vitro-cleavage assay could provide a rough assessment of gRNA efficiency, it largely excludes the interference of native genomic context. Transient in-vivo testing gives a proper assessment of the cleavage ability of editing reagents in a native genomic context. Here, we developed a modified protocol that offers highly efficient protoplast isolation from rice, <i>Arabidopsis,</i> and chickpea, using a sucrose gradient, transfection using PEG (polyethylene glycol), and validation of single guide RNAs (sgRNAs) cleavage efficiency of CRISPR-Cas9. We have optimized various parameters for PEG-mediated protoplast transfection and achieved high transfection efficiency using our protocol in both monocots and dicots. We introduced plasmid vectors containing Cas9 and sgRNAs targeting genes in rice, <i>Arabidopsis,</i> and chickpea protoplasts. Using dual sgRNAs, our CRISPR-deletion strategy offers straightforward detection of genome editing success by simple agarose gel electrophoresis. Sanger sequencing of PCR products confirmed the editing efficiency of specific sgRNAs. Notably, we demonstrated that isolated protoplasts can be stored for up to 24/48 h with little loss of viability, allowing a pause between isolation and transfection. This high-efficiency protocol for protoplast isolation and transfection enables rapid (less than 7 days) validation of sgRNA cleavage efficiency before proceeding with stable transformation. The isolation and transfection method can also be utilized for rapid validation of editing strategies, evaluating diverse editing reagents, regenerating plants from transfected protoplasts, gene expression studies, protein localization and functional analysis, and other applications.</p></div>","PeriodicalId":53135,"journal":{"name":"aBIOTECH","volume":null,"pages":null},"PeriodicalIF":4.6,"publicationDate":"2024-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s42994-024-00139-7.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140702659","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Zig, Zag, and ’Zyme: leveraging structural biology to engineer disease resistance","authors":"Alexander J. McClelland,&nbsp;Wenbo Ma","doi":"10.1007/s42994-024-00152-w","DOIUrl":"10.1007/s42994-024-00152-w","url":null,"abstract":"<div><p>Dynamic host–pathogen interactions determine whether disease will occur. Pathogen effector proteins are central players in such disease development. On one hand, they improve susceptibility by manipulating host targets; on the other hand, they can trigger immunity after recognition by host immune receptors. A major research direction in the study of molecular plant pathology is to understand effector-host interactions, which has informed the development and breeding of crops with enhanced disease resistance. Recent breakthroughs on experiment- and artificial intelligence-based structure analyses significantly accelerate the development of this research area. Importantly, the detailed molecular insight of effector–host interactions enables precise engineering to mitigate disease. Here, we highlight a recent study by Xiao et al., who describe the structure of an effector-receptor complex that consists of a fungal effector, with polygalacturonase (PG) activity, and a plant-derived polygalacturonase-inhibiting protein (PGIP). PGs weaken the plant cell wall and produce immune-suppressive oligogalacturonides (OGs) as a virulence mechanism; however, PGIPs directly bind to PGs and alter their enzymatic activity. When in a complex with PGIPs, PGs produce OG polymers with longer chains that can trigger immunity. Xiao et al. demonstrate that a PGIP creates a new active site tunnel, together with a PG, which favors the production of long-chain OGs. In this way, the PGIP essentially acts as both a PG receptor and enzymatic manipulator, converting virulence to defense activation. Taking a step forward, the authors used the PG-PGIP complex structure as a guide to generate PGIP variants with enhanced long-chain OG production, likely enabling further improved disease resistance. This study discovered a novel mechanism by which a plant receptor plays a dual role to activate immunity. It also demonstrates how fundamental knowledge, obtained through structural analyses, can be employed to guide the design of proteins with desired functions in agriculture.</p></div>","PeriodicalId":53135,"journal":{"name":"aBIOTECH","volume":null,"pages":null},"PeriodicalIF":4.6,"publicationDate":"2024-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s42994-024-00152-w.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140716615","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expanding the targeting scope of CRISPR/Cas9-mediated genome editing by Cas9 variants in Brassica","authors":"Wenjing Li,&nbsp;Xuan Li,&nbsp;Chunyang Wang,&nbsp;Guanzhong Huo,&nbsp;Xinru Zhang,&nbsp;Jintai Yu,&nbsp;Xiaoxiao Yu,&nbsp;Jing Li,&nbsp;Chao Zhang,&nbsp;Jianjun Zhao,&nbsp;Yan Li,&nbsp;Jun Li","doi":"10.1007/s42994-024-00155-7","DOIUrl":"10.1007/s42994-024-00155-7","url":null,"abstract":"<div><p>CRISPR/Cas9, presently the most widely used genome editing technology, has provided great potential for functional studies and plant breeding. However, the strict requirement for a protospacer adjacent motif (PAM) has hindered the application of the CRISPR/Cas9 system because the number of targetable genomic sites is limited. Recently, the engineered variants Cas9-NG, SpG, and SpRY, which recognize non-canonical PAMs, have been successfully tested in plants (mainly in rice, a monocot). In this study, we evaluated the targeted mutagenesis capabilities of these Cas9 variants in two important <i>Brassica</i> vegetables, Chinese cabbage (<i>Brassica rapa</i> spp. <i>pekinensis</i>) and cabbage (<i>Brassica oleracea</i> var. <i>capitata</i>). Both Cas9-NG and SpG induced efficient mutagenesis at NGN PAMs, while SpG outperformed Cas9-NG at NGC and NGT PAMs. SpRY achieved efficient editing at almost all PAMs (NRN &gt; NYN), albeit with some self-targeting activity at transfer (T)-DNA sequences. And SpRY-induced mutants were detected in cabbage plants in a PAM-less fashion. Moreover, an adenine base editor was developed using SpRY and TadA8e deaminase that induced A-to-G conversions within target sites using non-canonical PAMs. Together, the toolboxes developed here induced successful genome editing in Chinese cabbage and cabbage. Our work further expands the targeting scope of genome editing and paves the way for future basic research and genetic improvement in <i>Brassica</i>.</p></div>","PeriodicalId":53135,"journal":{"name":"aBIOTECH","volume":null,"pages":null},"PeriodicalIF":4.6,"publicationDate":"2024-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s42994-024-00155-7.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140737111","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Consumer transparency in the production chain for plant varieties produced using new genomic techniques","authors":"J. M. Lukasiewicz,&nbsp;C. C. M. van de Wiel,&nbsp;L. A. P. Lotz,&nbsp;M. J. M. Smulders","doi":"10.1007/s42994-024-00142-y","DOIUrl":"10.1007/s42994-024-00142-y","url":null,"abstract":"<div><p>Plants edited with new genomic techniques (NGTs) currently fall under the Genetically Modified Organisms Directive (2001/18/EC) in the European Union. In the proposal of the European Commission, NGT plants are partially exempted from the regulations of this directive. The proposal makes a distinction between two categories of NGT plants: NGT-1 and NGT-2. NGT-1 category plants are considered equal to plants obtained through conventional breeding methods. These plants will not be labelled for the consumer, although they will be labelled as seeds. NGT-2 category plants may be labelled with additional information as a positive incentive. Labelling of seeds of varieties made with gene editing, but not the products, would mean that most steps in the production chain are transparent, but not the last step towards consumers. The “right to know” and increasing knowledge of gene-edited food is a common theme in food labelling towards consumers. Here, we describe current labelling regimes and registers and how these may be applied to provide transparency on gene-edited products to consumers. Furthermore, we also look into consumer studies, which indicate a greater acceptance of gene-edited food among consumers, especially when additional benefits such as sustainability are mentioned.</p></div>","PeriodicalId":53135,"journal":{"name":"aBIOTECH","volume":null,"pages":null},"PeriodicalIF":4.6,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11224161/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141555971","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction: Fundamental and practical approaches for single-cell ATAC-seq analysis","authors":"Peiyu Shi,&nbsp;Yage Nie,&nbsp;Jiawen Yang,&nbsp;Weixing Zhang,&nbsp;Zhongjie Tang,&nbsp;Jin Xu","doi":"10.1007/s42994-024-00154-8","DOIUrl":"10.1007/s42994-024-00154-8","url":null,"abstract":"","PeriodicalId":53135,"journal":{"name":"aBIOTECH","volume":null,"pages":null},"PeriodicalIF":4.6,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s42994-024-00154-8.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140782399","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization and functional analysis of gerbera plant defensin (PDF) genes reveal the role of GhPDF2.4 in defense against the root rot pathogen Phytophthora cryptogea","authors":"Chunzhen Cheng,&nbsp;Huan Wu,&nbsp;Yongyan Zhang","doi":"10.1007/s42994-024-00146-8","DOIUrl":"10.1007/s42994-024-00146-8","url":null,"abstract":"<div><p>Gerbera (<i>Gerbera hybrida</i>), a major fresh cut flower crop, is very susceptible to root rot disease. Although plant defensins (PDFs), a major group of plant antimicrobial peptides, display broad-spectrum antifungal and antibacterial activities, <i>PDF</i> genes in gerbera have not been systematically characterized. Here, we identified and cloned nine <i>PDF</i> genes from gerbera and divided them into two classes based on phylogenetic analysis. Most Class I <i>GhPDF</i> genes were highly expressed in petioles, whereas all Class II <i>GhPDF</i> genes were highly expressed in roots. <i>Phytophthora cryptogea</i> inoculation strongly upregulated all Class II <i>GhPDF</i> genes in roots and upregulated all Class I <i>GhPDF</i> genes in petioles. Transient overexpression of <i>GhPDF1.5</i> and <i>GhPDF2.4</i> inhibited <i>P. cryptogea</i> infection in tobacco (<i>Nicotiana benthamiana</i>) leaves. Transient overexpression of <i>GhPDF2.4</i>, but not <i>GhPDF1.5</i>, significantly upregulated <i>ACO</i> and <i>LOX</i> gene expression in tobacco leaves, indicating that overexpressing <i>GhPDF2.4</i> activated the jasmonic acid/ethylene defense pathway and that the two types of GhPDFs have different modes of action. Prokaryotically expressed recombinant GhPDF2.4 inhibited mycelial growth and delayed the hyphal swelling of <i>P. cryptogea</i>, in vitro, indicating that GhPDF2.4 is a morphogenetic defensin. Moreover, the addition of GhPDF2.4 to plant culture medium alleviated the root rot symptoms of in vitro<i>-</i>grown gerbera seedlings and greatly reduced pathogen titer in <i>P. cryptogea</i>-inoculated gerbera roots in the early stages of treatment. Our study provides a basis for the use of GhPDFs, especially GhPDF2.4, for controlling root rot disease in gerbera.</p></div>","PeriodicalId":53135,"journal":{"name":"aBIOTECH","volume":null,"pages":null},"PeriodicalIF":4.6,"publicationDate":"2024-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s42994-024-00146-8.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140360115","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Removal of the C4-domain preserves the drought tolerance enhanced by CsMYB4a and eliminates the negative impact of this transcription factor on plant growth","authors":"Mingzhuo Li,&nbsp;Guoliang Ma,&nbsp;Xiu Li,&nbsp;Lili Guo,&nbsp;Yanzhi Li,&nbsp;Yajun Liu,&nbsp;Wenzhao Wang,&nbsp;Xiaolan Jiang,&nbsp;De-Yu Xie,&nbsp;Liping Gao,&nbsp;Tao Xia","doi":"10.1007/s42994-024-00149-5","DOIUrl":"10.1007/s42994-024-00149-5","url":null,"abstract":"<div><p>The MYB4 transcription factor family regulates plant traits. However, their overexpression often results in undesirable side effects like growth reduction. We have reported a green tea (<i>Camellia sinensis</i>) MYB4 transcription factor (CsMYB4) that represses the phenylpropanoid and shikimate pathways and stunts plant growth and development. In the current study, we observed that in <i>CsMYB4a</i> transgenic tobacco (<i>Nicotiana tabacum</i>) plants, primary metabolism was altered, including sugar and amino acid metabolism, which demonstrated a pleiotropic regulation by CsMYB4a. The <i>CsMYB4a</i> transgenic tobacco plants had improved drought tolerance, which correlated to alterations in carbohydrate metabolism and an increase in proline content, as revealed by metabolic profiling and transcriptomic analysis. To mitigate the undesirable repressive side effects on plant traits, including dwarfism, shrunken leaves, and shorter roots of <i>CsMYB4a</i> transgenic plants, we deleted the C4 domain of CsMYB4a to obtain a CsMYB4a-DC4 variant and then overexpressed it in transgenic plants (CsMYB4a-DC4). These CsMYB4a-DC4 plants displayed a normal growth and had improved drought tolerance. Metabolite analysis demonstrated that the contents of carbohydrates and proline were increased in these transgenic plants. Our findings suggest that  an approriate modification of TFs can generate novel crop traits, thus providing potential agricultural benefits and expanding its application to various crops.</p></div>","PeriodicalId":53135,"journal":{"name":"aBIOTECH","volume":null,"pages":null},"PeriodicalIF":4.6,"publicationDate":"2024-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s42994-024-00149-5.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140369003","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced editing efficiency in Arabidopsis with a LbCas12a variant harboring D156R and E795L mutations","authors":"Cuiping Xin,&nbsp;Dexin Qiao,&nbsp;Junya Wang,&nbsp;Wei Sun,&nbsp;Zhenghong Cao,&nbsp;Yu Lu,&nbsp;Yuanyuan Jiang,&nbsp;Yiping Chai,&nbsp;Xue-Chen Wang,&nbsp;Qi-jun Chen","doi":"10.1007/s42994-024-00144-w","DOIUrl":"10.1007/s42994-024-00144-w","url":null,"abstract":"<div><p>Cas12a (Cpf1), a Class 2 Type V CRISPR/Cas nuclease, has several unique attributes for genome editing and may provide a valuable alternative to Cas9. However, a low editing efficiency due to temperature sensitivity and insufficient cleavage activity of the Cas12a nuclease are major obstacles to its broad application. In this report, we generated two variants, ttAsCas12 Ultra and ttLbCas12a Ultra harboring three (E174R, M537R, and F870L) or two (D156R and E795L) mutations, respectively, by combining the mutations from the temperature-tolerant variants ttAsCas12a (E174R) and ttLbCas12a (D156R), and those from the highly active variants AsCas12a Ultra (M537R and F870L) and LbCas12a Ultra (E795L). We compared editing efficiencies of the five resulting Cas12a variants (LbCas12a, ttLbCas12a, ttLbCas12a Ultra, AsCas12a Ultra, and ttAsCas12 Ultra) at six target sites of four genes in Arabidopsis (<i>Arabidopsis thaliana</i>). The variant ttLbCas12a Ultra, harboring the D156R and E795L mutations, exhibited the highest editing efficiency of all variants tested in Arabidopsis and can be used to generate homozygous or biallelic mutants in a single generation in Arabidopsis plants grown at 22 °C. In addition, optimization of ttLbCas12a Ultra, by varying nuclear localization signal sequences and codon usage, further greatly improved editing efficiency. Collectively, our results indicate that ttLbCas12a Ultra is a valuable alternative to Cas9 for editing genes or promoters in Arabidopsis.</p></div>","PeriodicalId":53135,"journal":{"name":"aBIOTECH","volume":null,"pages":null},"PeriodicalIF":4.6,"publicationDate":"2024-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s42994-024-00144-w.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140379325","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}