Programmed gRNA Removal System for CRISPR-Cas9-Mediated Multi-Round Genome Editing in Bacillus subtilis

CRISPR/Cas9 has become a simple and powerful genome editing tool for many organisms. However, multi-round genome editing should replace single-guide RNA (sgRNA) every round, which is laborious and time-consuming. Here, we have developed a multi-round genome editing system in which genome editing and the programmed removal of the sgRNA have sequentially occurred in a growth-dependent manner in Bacillus subtilis. The system contains two plasmids, one containing a cas9 gene and the other containing two sgRNAs and a donor DNA for homology directed repair (HDR). The two sgRNAs are chromosome-targeting (sgRNAct) and self-targeting (sgRNAst) under the control of a constitutive promoter and sporulation-specific promoter, respectively. In the growth phase, the sgRNAct is transcribed and complexed with the Cas9 to edit the chromosomal target, while the sgRNAst is transcribed in the sporulation phase and complexed with the Cas9 to attack its own plasmid. Therefore, the system automatically makes the cell ready for next-round genome editing during cultivation. The system was approved through the sequential deletion of eight extracellular protease genes in the B. subtilis, suggesting that it can be used for versatile applications in multi-round genome editing

Investigating key attributes in experience and satisfaction of hotel customer using online review data

© 2019 by the authors. Licensee MDPI, Basel, Switzerland. With the development of social media, customers are sharing their experiences, and it is rapidly spreading as a form of online review. That is why the online review has become a significant information source affecting customers\u27 purchase intention and behavior. Therefore, it is important to understand the customer\u27s experience shown in the online review in order to maintain sustainable customer satisfaction and loyalty. The purpose of this study is to investigate what are the key attributes and the structural relationship of those key attributes. To accomplish this purpose, a total of 6596 hotel reviews were collected from Google (google.com). A frequency analysis using text mining was performed to figure out the most frequently mentioned attributes. In addition, semantic network analysis, factor analysis, and regression analysis were applied to understand the experience and satisfaction of the hotel customer. As a result, the top 99 keywords were divided into four groups such as Intangible Service , Physical Environment , Purpose , and Location . The factor analysis reduced the dimension of the original 64 keywords to 22 keywords, and grouped them into five factors, which are Access , F&B (Food and Beverage) , Purpose , Tangibles , and Empathy . Based on these results, theoretical and practical implications for sustainable hotel marketing strategies are suggested

Factors affecting customer intention to use online food delivery services before and during the COVID-19 pandemic

With the emerging popularity of online food delivery (OFD) services, this research examined predictors affecting customer intention to use OFD services amid the Coronavirus disease (COVID-19) pandemic. Specifically, Study 1 examined the moderating effect of the pandemic on the relationship between six predictors (perceived usefulness, perceived ease of use, price saving benefit, time saving benefit, food safety risk perception, and trust) and OFD usage intention, and Study 2 extended the model by adding customer perceptions of COVID-19 (perceived severity and vulnerability) during the pandemic. Study 1 showed that all of the predictors except food safety risk perception significantly affected OFD usage intention, but no moderation effect of COVID-19 was found. In Study 2, while perceived severity and vulnerability had no significant impact on OFD usage intention, the altered effects of socio-demographic variables during the COVID-19 pandemic were found. Theoretical and managerial implications are provided

A Novel Function Complexity-Based Code Migration Policy for Reducing Power Consumption

Embedded system designs have changed greatly owing to rapid developments in both hardware and software technology. Typical design should consider hardware limitations, such as size, weight, or battery capacity. In other words, the designs are heavily dependent on the hardware component. Since hardware can deteriorate and degenerate, hardware-aware software design is needed to achieve power-efficient embedded systems. Studies usually focus on the microprocessor in terms of optimizing power consumption. Besides computation, however, the system also consumes power when executing programs. A lot of memory accesses result in the entire execution, it should be considered to minimize for more efficient designs. Modern embedded systems often use heterogeneous memory to benefit from different characteristics of memory devices. This study aims to optimize the power efficiency of heterogeneous memory in embedded systems. We have proposed a detailed function complexity concept to identify specific function units in a program that consume less power in migrated memory. Using the function complexity, function selection algorithm is proposed to select a unique function which improves most after the migration. Experiments and quantitative analyses with various benchmarks have been performed to prove the validity of the proposed algorithm. Power consumption is successfully minimized by migrating certain function of a program in low-power memory

Identification of RipAZ1 as an avirulence determinant of Ralstonia solanacearum in Solanum americanum

Ralstonia solanacearum causes bacterial wilt disease in many plant species. Type III-secreted effectors (T3Es) play crucial roles in bacterial pathogenesis. However, some T3Es are recognized by corresponding disease resistance proteins and activate plant immunity. In this study, we identified the R. solanacearum T3E protein RipAZ1 (Ralstonia injected protein AZ1) as an avirulence determinant in the black nightshade species Solanum americanum. Based on the S. americanum accession-specific avirulence phenotype of R. solanacearum strain Pe_26, 12 candidate avirulence T3Es were selected for further analysis. Among these candidates, only RipAZ1 induced a cell death response when transiently expressed in a bacterial wilt-resistant S. americanum accession. Furthermore, loss of ripAZ1 in the avirulent R. solanacearum strain Pe_26 resulted in acquired virulence. Our analysis of the natural sequence and functional variation of RipAZ1 demonstrated that the naturally occurring C-terminal truncation results in loss of RipAZ1-triggered cell death. We also show that the 213 amino acid central region of RipAZ1 is sufficient to induce cell death in S. americanum. Finally, we show that RipAZ1 may activate defence in host cell cytoplasm. Taken together, our data indicate that the nucleocytoplasmic T3E RipAZ1 confers R. solanacearum avirulence in S. americanum. Few avirulence genes are known in vascular bacterial phytopathogens and ripAZ1 is the first one in R. solanacearum that is recognized in black nightshades. This work thus opens the way for the identification of disease resistance genes responsible for the specific recognition of RipAZ1, which can be a source of resistance against the devastating bacterial wilt disease

Modeling the Power Consumption of Function-Level Code Relocation for Low-Power Embedded Systems

The problems associated with the battery life of embedded systems were addressed by focusing on memory components that are heterogeneous and are known to meaningfully affect the power consumption and have not been fully exploited thus far. Our study establishes a model that predicts and orders the efficiency of function-level code relocation. This is based on extensive code profiling that was performed on an actual system to discover the impact and was achieved by using function-level code relocation between the different types of memory, i.e., flash memory and static RAM, to reduce the power consumption. This was accomplished by grouping the assembly instructions to evaluate the distinctive power reduction efficiency depending on function code placement. As a result of the profiling, the efficiency of the function-level code relocation was the lowest at 11.517% for the branch and control groups and the highest at 12.623% for the data processing group. Further, we propose a prior relocation-scoring model to estimate the effective relocation order among functions in a program. To demonstrate the effectiveness of the proposed model, benchmarks in the MiBench benchmark suite were selected as case studies. The experimental results are consistent in terms of the scored outputs produced by the proposed model and measured power reduction efficiencies

Implementasi Hidden Curriculum dalam pembentukan karakter Religius peserta didik: Studi Multi Situs di MAN Model dan SMA Muhammadiyah Al-Amin di Sorong

مستخلص البحث تشكيل الطابع الشخصي في التعليم الرسمي لا يمكن فصلها عن إشراك مدير المدرسة، والمعلمين وأولياء الأمور الذين ساهموا في تحديد نجاحه بشكل كبير. وبالإضافة إلى ذلك، هناك بعض العناصر المخفية الأخرى من عناصر المنهج المدرسي الرسمي. المنهج الدراسي الخفي هو واحد من الجهود التي غالبا ما يتم تركها في تشكيل الطابع الشخصي. مثل، إدارة أنشطة التعليم والتعلم، والأنشطة اللاصفية، وتكوين جو التعلم وبيئة المدرسة المتميزة، والتعويد، وغرس القيم والأخلاق الحسنة لدعم نجاح عملية تشكيل الطابع الشخصي. يهدف هذا البحث إلى الكشف عن تنفيذ المنهج الدراسي الخفي في تشكيل الطابع الديني للطلبة في المدرسة الثانوية الدينية الحكومية النموذجية والمدرسة الثانوية العامة المحمدية "الأمين" سورونج، ويركز على ما يلي: (1) شكل التنفيذ، (2) تنفيذ الاستراتيجية، و(3) الأثار المترتبة من تنفيذ المنهج الدراسي الخفي في تشكيل الطابع الديني للطلبة. استخدم هذا البحث المدخل النوعي، ومن نوع البحث الميداني ومع تصميم دراسة متعدد المواقع. ويتم جمع البيانات من خلال المراقبة على المشاركين، والمقابلة المتعمقة، والوثائق. وتشمل طريقة تحليل البيانات تحديد البيانات، وعرضها، والاستنتاج منها أو التحقق من صحتها. وأمّا التحقق من صحة البيانات فيتم عن طريق تمديد المراقبة، وزيادة القدرة على التحمل، التثليث، وتحليل الحالة السلبية، والتحقق. أظهرت نتائج هذا البحث ما يلي: (1) يحتوي شكل تنفيذ المنهج الدراسي الخفي في تشكيل الطابع الديني للطلبة في المدرسة الثانوية الدينية الحكومية النموذجية والمدرسة الثانوية العامة المحمدية "الأمين" سورونج على الجانب الهيكلي والجانب الثقافي ويتم تنفيذهما داخل الفصل الدراسي أو خارجه، (2) تشمل استراتيجية تنفيذ المنهج الدراسي الخفي في تشكيل الطابع الديني للطلبة في المدرسة الثانوية الدينية الحكومية النموذجية والمدرسة الثانوية العامة المحمدية "الأمين" سورونج جميع الأعمال التي يقوم بها أصحاب المصلحة التعليمية، و(3) الأثار المترتبة من تنفيذ المنهج الدراسي الخفي في تشكيل الطابع الديني للطلبة في المدرسة الثانوية الدينية الحكومية النموذجية والمدرسة الثانوية العامة المحمدية "الأمين" سورونج تشمل قيمة العقيدة، والعبادة، والأخلاق. ABSTRACT Character building in formal education cannot be separated from the involvement of principals, teachers, and parents who have a great role in determining its success. In addition, there are some hidden elements beside the formal school’s curriculum. The Hidden Curriculum is one of the efforts that are often ignored in character building. Those are the management of teaching and learning activities, extracurricular activities, the creation of an atmosphere of learning and characterized school environment, habituation, and embedment of good values and ethics to support the success of the character formation process. This study aims to reveal the implementation of hidden curriculum in building religious character of the students in MAN Model and SMA Muhammadiyah Al-Amin in Sorong, with sub focuses of study include: (1) implementation model, (2) implementation efforts, and (3) the implementation impact of the hidden curriculum in building the religious character of the students. This study employs a qualitative approach using field research and multi-site study design. The data collection is performed through participant observation, in-depth interviews, and documentation. The data analysis techniques include data reduction, data presentation, and conclusion drawing or verification. The data validity is checked through extension of observation, increased persistence, triangulation, negative case analysis, and member check. The results showed that: (1) the hidden curriculum implementation model in the building students’ religious character in MAN Model and SMA Muhammadiyah Al-Amin in Sorong covers structural and cultural aspects implemented inside and outside the classroom, (2) the hidden curriculum implementation efforts in building the students’ religious character in MAN Model and SMA Muhammadiyah Al-Amin in Sorong cover all efforts that performed by all components of educationional stakeholders, and (3) the impacts of the hidden curriculum implementation in building the students’ religious character in MAN Model and SMA Muhammadiyah Al-Amin in Sorong cover the value of aqeedah, worship, and morals ABSTRAK Pembentukan karakter dalam pendidikan formal tidak dapat terlepas dari keterlibatan kepala sekolah, guru, dan orang tua siswa yang memiliki andil sangat besar dalam menentukan keberhasilannya. Selain itu, yang tak kalah pentingnya, terdapat beberapa unsur yang tersembunyi selain unsur kurikulum formal sekolah. The Hidden Curriculum adalah salah satu upaya yang sering terabaikan dalam pembentukan karakter. Seperti, pengelolaan kegiatan belajar mengajar, kegiatan ekstrakurikuler, penciptaan suasana belajar dan lingkungan sekolah berkarakter, pembiasaan, dan pembudayaan nilai dan etika yang baik dapat mendukung keberhasilan proses pembentukan karakter. Penelitian ini bertujuan untuk mengungkapkan implementasi hidden curriculum dalam pembentukan karakter religius peserta didik di MAN Model dan SMA Muhammadiyah Al-Amin di Sorong, dengan sub fokus penelitian meliputi: (1) bentuk pelaksanaan, (2) upaya pelaksanaan, dan (3) dampak dari pelaksanaan hidden curriculum dalam pembentukan karakter religius peserta didik. Penelitian ini menggunakan pendekatan penelitian kualitatif dengan jenis penelitian lapangan dan dengan rancangan studi multi situs. Pengumpulan data dilakukan dengan pengamatan terlibat, wawancara mendalam, dan dokumentasi. Teknik analisis data meliputi reduksi data, penyajian data, dan pemberian kesimpulan atau verifikasi. Pengecekan keabsahan data dilakukan dengan cara perpanjangan pengamatan, peningkatan ketekunan, triangulasi, analisis kasus negatif, dan member check. Hasil penelitian menunjukkan bahwa: (1) bentuk pelaksanaan hidden curriculum dalam pembentukan karakter religius peserta didik di MAN Model dan SMA Muhammadiyah Al-Amin di Sorong mencakup aspek struktural dan kultural yang pelaksanaannya di dalam dan di luar kelas, (2) upaya pelaksanaan hidden curriculum dalam pembentukan karakter religius peserta didik di MAN Model dan SMA Muhammadiyah Al-Amin di Sorong meliputi seluruh usaha yang dilakukan seluruh komponen stakeholders pendidikan, dan (3) dampak pelaksanaan hidden curriculum dalam pembentukan karakter religius peserta didik di MAN Model dan SMA Muhammadiyah Al-Amin di Sorong meliputi nilai aqidah, ibadah, dan akhlak

The Combined Role of Independence in Self-Concept and A Collectivistic Value Orientation in Group-Focused Enmity in Korea

Previous research (Zick et al. 2008) suggested that animosity toward social minorities in Europe is intertwined, forming a syndrome of Group-Focused Enmity (GFE). In the current research, we extended the notion of GFE by identifying the GFE structure in a non-European context (South Korea). We also tested a novel hypothesis that stipulates an interplay between individuals’ self-concept and their value orientation in predicting the overall level of GFE. Two nation-wide surveys in South Korea showed that antagonism toward social minorities that have typically been marginalized and devalued in that country forms GFE while reflecting the unique intergroup context of Korean society. Further, we found as expected that independence in self-concept and a collectivistic value orientation jointly predict lower levels of GFE (Study 1). When political orientation and national identification were taken into account, the predicted interaction was observed only on antagonism toward ingroup minorities but not toward outgroup minorities by race or ethnicity (Study 2). Implications of these findings and directions for research on GFE are discussed

Biosensors Based on Bivalent and Multivalent Recognition by Nucleic Acid Scaffolds

Several biological macromolecules adopt bivalent or multivalent interactions to perform various cellular processes. In this regard, the development of molecular constructs presenting multiple ligands in a specific manner is becoming crucial for the understanding of multivalent interactions and for the detection of target macromolecules. Nucleic acids are attractive molecules to achieve this goal because they are capable of forming various, structurally well-defined 2D or 3D nanostructures and can bear multiple ligands on their structures with precisely controlled ligand–ligand distances. Thanks to the features of nucleic acids, researchers have proposed a wide range of bivalent and multivalent binding agents that strongly bind to target biomolecules; consequently, these findings have uncovered new biosensing strategies for biomolecule detection. To date, various bivalent and multivalent interactions of nucleic acid architectures have been applied to the design of biosensors with enhanced sensitivity and target accuracy. In this review, we describe not only basic biosensor designs but also recently designed biosensors operating through the bivalent and multivalent recognition of nucleic acid scaffolds. Based on these designs, strategies to transduce bi- or multivalent interaction signals into readable signals are discussed in detail, and the future prospects and challenges of the field of multivalence-based biosensors are explored

Biosensors Based on Bivalent and Multivalent Recognition by Nucleic Acid Scaffolds

Several biological macromolecules adopt bivalent or multivalent interactions to perform various cellular processes. In this regard, the development of molecular constructs presenting multiple ligands in a specific manner is becoming crucial for the understanding of multivalent interactions and for the detection of target macromolecules. Nucleic acids are attractive molecules to achieve this goal because they are capable of forming various, structurally well-defined 2D or 3D nanostructures and can bear multiple ligands on their structures with precisely controlled ligand–ligand distances. Thanks to the features of nucleic acids, researchers have proposed a wide range of bivalent and multivalent binding agents that strongly bind to target biomolecules; consequently, these findings have uncovered new biosensing strategies for biomolecule detection. To date, various bivalent and multivalent interactions of nucleic acid architectures have been applied to the design of biosensors with enhanced sensitivity and target accuracy. In this review, we describe not only basic biosensor designs but also recently designed biosensors operating through the bivalent and multivalent recognition of nucleic acid scaffolds. Based on these designs, strategies to transduce bi- or multivalent interaction signals into readable signals are discussed in detail, and the future prospects and challenges of the field of multivalence-based biosensors are explored