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Abstract Huntington’s disease (HD) is a severe inherited neurological disorder caused by a CAG repeat expansion in the huntingtin gene (HTT), leading to the accumulation of mutant huntingtin with polyglutamine repeats. Despite its severity, there is no cure for this debilitating disease. HTT lowering strategies, including antisense oligonucleotides (ASO) showed promising results very recently. Attempts to develop stem cell-based therapeutics have shown efficacy in preclinical HD models. Using an HD patient’s autologous cells, which have genetic defects, may hamper therapeutic efficacy due to mutant HTT. Pretreating these cells to reduce mutant HTT expression and transcription may improve the transplanted cells’ therapeutic efficacy. To investigate this, we targeted the SUPT4H1 gene that selectively supports the transcription of long trinucleotide repeats. Transplanting SUPT4H1-edited HD-induced pluripotent stem cell-derived neural precursor cells (iPSC-NPCs) into the YAC128 HD transgenic mouse model improved motor function compared to unedited HD iPSC-NPCs. Immunohistochemical analysis revealed reduced mutant HTT expression without compensating wild-type HTT expression. Further, SUPT4H1 editing increased neuronal and decreased reactive astrocyte differentiation in HD iPSC-NPCs compared to the unedited HD iPSC-NPCs. This suggests that ex vivo editing of SUPT4H1 can reduce mutant HTT expression and provide a therapeutic gene editing strategy for autologous stem cell transplantation in HD.

The Streptococcus pyogenes CRISPR/Cas9 (SpCas9) system is now widely utilized to generate genome engineered mice; however, some studies raised issues related to off-target mutations with this system. Herein, we utilized the Campylobacter jejuni Cas9 (CjCas9) system to generate knockout mice. We designed sgRNAs targeting mouse Tyr or Foxn1 and microinjected into zygotes along with CjCas9 mRNA. We obtained newborn mice from the microinjected embryos and confirmed that 50% (Tyr) and 38.5% (Foxn1) of the newborn mice have biallelic mutation on the intended target sequences, indicating efficient genome targeting by CjCas9. In addition, we analyzed off-target mutations in founder mutant mice by targeted deep sequencing and whole genome sequencing. Both analyses revealed no off-target mutations at potential off-target sites predicted in silico and no unexpected random mutations in analyzed founder animals. In conclusion, the CjCas9 system can be utilized to generate genome edited mice in a precise manner.

Genome editing: Improving efficiency by improving DNA repair A method to improve the efficiency of repairing double-stranded DNA breaks facilitates the production of animals with edited genomes. The CRISPR/Cas9 system has been revolutionising biomedical research as it allows the insertion or deletion of any DNA sequence at specific sites by making double-strand DNA breaks. However, the insertion of precise genetic changes is limited by the relatively low efficiency of the repair mechanism, which requires a template DNA molecule to promote error-free repair of the DNA strands. A team of South Korean scientists led by Su Cheong Yeom at Seoul National University and Han Woong Lee at Yonsei University show that delivering multiple overlapping single-guide RNAs (that share at least five base pairs of the target sequence) as a DNA template significantly improves the repair and thus the production of mice with a targeted gene insertion.

KO mice provide an excellent tool to determine roles of specific genes in biomedical filed. Traditionally, knockout mice were generated by homologous recombination in embryonic stem cells. Recently, engineered nucleases, such as zinc finger nuclease, transcription activator-like effector nuclease and clustered regularly interspaced short palindromic repeats (CRISPR), were used to produce knockout mice. This new technology is useful because of high efficiency and ability to generate biallelic mutation in founder mice. Until now, most of knockout mice produced using engineered nucleases were C57BL/6 strain. In the present study we used CRISPR-Cas9 system to generate knockout mice in FVB strain. We designed and synthesized single guide RNA (sgRNA) of CRISPR system for targeting gene, Abtb2. Mouse zygote were obtained from superovulated FVB female mice at 8-10 weeks of age. The sgRNA was injected into pronuclear of the mouse zygote with recombinant Cas9 protein. The microinjected zygotes were cultured for an additional day and only cleaved embryos were selected. The selected embryos were surgically transferred to oviduct of surrogate mother and offsprings were obtained. Genomic DNA were isolated from the offsprings and the target sequence was amplified using PCR. In T7E1 assay, 46.7% among the offsprings were founded as mutants. The PCR products were purified and sequences were analyzed. Most of the mutations were founded as deletion of few sequences at the target site, however, not identical among the each offspring. In conclusion, we found that CRISPR system is very efficient to generate knockout mice in FVB strain.

In vitro culture of murine embryos is an important step for in vitro production systems including in vitro fertilization and generations of genetically engineered mice. M16 is widely used commercialized culture media for the murine embryos. Compared to other media such as potassium simplex optimization medium, commercial M16 (Sigma) media lacks of amino acid, glutamine and antibiotics. In the present study, we optimized M16 based embryo culture system using commercialized antibiotics-glutamine or amino acids supplements. In vivo derived murine zygote were M16 media were supplemented with commercial Penicillin-Streptomycin-Glutamine solution (PSG; Gibco) or MEM Non- Essential Amino Acids solution (NEAA; Gibco) as experimental design. Addition of PSG did not improved cleavage and blastocyst rates. On the other hand, cleavage rate is not different between control and NEAA treated group, however, blastocyst formation is significantly (P

In ancient history, individual lives paralleled nations in their rise and fall, thereby reflecting their destiny; however, individuals were overshadowed by the glorified history of a collective entity. Therefore, familial or tribal traditions reflected in genealogies sometimes contradicted official history; a good example in this regard is 1 Chronicles 7:20–27. An initial reading of the genealogy contained therein focused on its literary and rhetorical implications; subsequently, its homiletical implications were extended. From a literary perspective, the ending of Ephraim’s genealogy with Joshua was the Chronicler’s special device that placed the first unsuccessful exploitation by Ephraim’s sons as an overture to the long history of conquest that followed. The scriptural text contextualised Joshua’s positive judgement regarding the Promised Land and his election as Moses’ successor. From a homiletical perspective, Ephraim’s genealogy generated insights about failure and tragedy and hope for the fulfilment of God’s promise, also likening the life of faith to a journey of perseverance. Research findings revealed similarities in the literary and homiletic meaning of Ephraim’s genealogy with that of Terah in Genesis 11:27–32. Intradisciplinary and/or interdisciplinary implications: Homileticians used to complain that biblical studies were more oriented towards historic-critical interest than towards preaching. Results of this research, which relate to the discipline of Old Testament Studies, show how a genealogical text can be relevant for homiletic and pastoral use in church ministry.

The reason underlying God's divergent actions towards the offerings of Cain and Abel has been highly disputed. It has been often assumed that the reason mainly lies in the two offerers' differing dispositions and attitudes or in the quality and character of their offerings. However, the close relationship between YHWH and Cain and the differences between the text's descriptions of Cain and of Abel provide a more concrete rationale for God's action: Through the seemingly unjust rejection of Cain's offering, YHWH calls on him to recognize his relationship and responsibility for his inferior brother. That the relation to others is inseparable from the relation to God becomes evident once more in the punishment. [ABSTRACT FROM AUTHOR]

The article offers information on the workshop "Terminology Design: Quality Criteria and Evaluation Methods" held in Genoa, Italy on May 28, 2006. The workshop highlights various researches on the topics of terminology evaluation and multilingualism. Chairperson of the workshop, Fidéla Ibekwe-SanJuan, did the opening with the introduction of the workshop process and presenting its major themes primarily regarding on the three sub-topics of multilingual terminology evaluation such as corpora, tools and ontologies. Among the presenters during the workshop include Barbara Inge Karsch, Jorge Vivaldi and Horacio Rodriguez.

In terminology, the predominance of nouns is an incontestable phenomenon. In Korean terminologies, this predominance of nouns is even more notable because the meaning and function associated with adjectives in Indo-European languages are often realized in noun form. However, the rarer adjectival terms are, the more they are used in restrictive, repetitive ways in specialized domains. Thus, it is important to distinguish the different senses of these terms. In this work, focusing on semantic characterization in terminology, we distinguish the different senses of adjectival medical terms by applying lexico-semantic criteria (L’Homme 2004a) and by classifying the arguments of the adjective into semantic categories (Bae et al. 2002). With this work, we aim to enrich terminological descriptions found in Korean medical dictionnaries by demonstrating empirically a method for distinguishing the different senses of adjectival medical terms. To achieve our goal, we used the KAIST corpus, composed of medical texts (1,500,000 eojeols), and a group of texts on various subjects (40,000,000 eojeols). [ABSTRACT FROM AUTHOR]