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Utilizing cell-free DNA to validate targeted disruption of MYO7A in rhesus macaque pre-implantation embryos

  • Junghyun Ryu;Fernanda C. Burch;Emily Mishler;Martha Neuringer;Jon D. Hennebold;Carol Hanna
    • 한국동물생명공학회지
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    • 제37권4호
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    • pp.292-297
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    • 2022
  • Direct injection of CRISPR/Cas9 into zygotes enables the production of genetically modified nonhuman primates (NHPs) essential for modeling specific human diseases, such as Usher syndrome, and for developing novel therapeutic strategies. Usher syndrome is a rare genetic disease that causes loss of hearing, retinal degeneration, and problems with balance, and is attributed to a mutation in MYO7A, a gene that encodes an uncommon myosin motor protein expressed in the inner ear and retinal photoreceptors. To produce an Usher syndrome type 1B (USH1B) rhesus macaque model, we disrupted the MYO7A gene in developing zygotes. Identification of appropriately edited MYO7A embryos for knockout embryo transfer requires sequence analysis of material recovered from a trophectoderm (TE) cell biopsy. However, the TE biopsy procedure is labor intensive and could adversely impact embryo development. Recent studies have reported using cell-free DNA (cfDNA) from embryo culture media to detect aneuploid embryos in human in vitro fertilization (IVF) clinics. The cfDNA is released from the embryo during cell division or cell death, suggesting that cfDNA may be a viable resource for sequence analysis. Moreover, cfDNA collection is not invasive to the embryo and does not require special tools or expertise. We hypothesized that selection of appropriate edited embryos could be performed by analyzing cfDNA for MYO7A editing in embryo culture medium, and that this method would be advantageous for the subsequent generation of genetically modified NHPs. The purpose of this experiment is to determine whether cfDNA can be used to identify the target gene mutation of CRISPR/Cas9 injected embryos. In this study, we were able to obtain and utilize cfDNA to confirm the mutagenesis of MYO7A, but the method will require further optimization to obtain better accuracy before it can replace the TE biopsy approach.

Bacillus turingiensis 변종(變種)들로부터의 Plasmid DNA 추출(抽出) 및 분리(分離) (A Rapid Procedure for Screening and Isolation of Various Sizes of Plasmid DNA in Serovars of Bacillus thuringiensis)

  • 이영근;로베트 엠 파우스트;강석권;페트리시아 이 멕콜리;케롤 엘 메이어-다운링
    • 한국응용곤충학회지
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    • 제24권1호
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    • pp.45-50
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    • 1985
  • Bacillus thuringiensis 변종(變種)들로부터 Extrachromosomal DNA를 추출분리(抽出分離)코저 종래(從來)의 방법(方法)을 보완(補完)하여 적용(適用)한바 분자량(分子量)의 크기가 1 Megadalton에서 135 Megadalton에 이르는 plasmid들을 분리(分離)함에 보다 효과적(效果的)이었고 또 이 plasmid들을 이용(利用), 제한효소(制限酵素)에 의(依)한 유전자배열작성(遺傳子配列作成) 및 gene Cloning을 하는데 비교적(比較的) 안정(安定)된 많은 양(量)의 세포용해물(細胞溶解物)을 얻을 수 있었다. 파리목과 나비목에 각기(各其) 독성(毒性)이 다른 Bacillus thuringiensis 6개(個) 변종(變種)으로부터 plasmid들을 분리(分離)한 결과(結果) 분자량(分子量)이 큰 50 Megadalton 이상(以上)의 plasmid들이 공시(供試) 된 모든 변종(變種)으로부터 추출(抽出)되었으며 이들 plasmid의 수(數)를 보면 israelensis로부터 8개(個) kurstaki로부터 10개(個) $aizawa{\ddot{u}}$로부터 13개(個) dendrolimus로부터 2개(個), finitimus로부터 1개(個) 그리고 yunnanensis로부터 6개(個)가 각각(各各) 검출(檢出)되었다. 공시(供試)된 변종중(變種中) 4개(個)의 변종(變種)으로부터는 2 Megadalton 이하(以下)의 적은 plasmid들도 추출(抽出)되었다.

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The Philippines Coconut Genomics Initiatives: Updates and Opportunities for Capacity Building and Genomics Research Collaboration

  • Hayde Flandez-Galvez;Darlon V. Lantican;Anand Noel C. Manohar;Maria Luz J. Sison;Roanne R. Gardoce;Barbara L. Caoili;Alma O. Canama-Salinas;Melvin P. Dancel;Romnick A. Latina;Cris Q. Cortaga;Don Serville R. Reynoso;Michelle S. Guerrero;Susan M. Rivera;Ernesto E. Emmanuel;Cristeta Cueto;Consorcia E. Reano;Ramon L. Rivera;Don Emanuel M. Cardona;Edward Cedrick J. Fernandez ;Robert Patrick M. Cabangbang;Maria Salve C. Vasquez;Jomari C. Domingo;Reina Esther S. Caro;Alissa Carol M. Ibarra;Frenzee Kroeizha L. Pammit;Jen Daine L. Nocum;Angelica Kate G. Gumpal;Jesmar Cagayan;Ronilo M. Bajaro;Joseph P. Lagman;Cynthia R. Gulay;Noe Fernandez-Pozo;Susan R. Strickler;Lukas A. Mueller
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2022년도 추계학술대회
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    • pp.30-30
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    • 2022
  • Philippines is the second world supplier of coconut by-products. As its first major genomics project, the Philippine Genome Center program for Agriculture (PGC-Agriculture) took the challenge to sequence and assemble the whole coconut genome. The project aims to provide advance genetics tools for our collaborating coconut researchers while taking the opportunity to initiate local capacity. Combination of different NGS platforms was explored and the Philippine 'Catigan Green Dwarf' (CATD) variety was selected with the breeders to be the crop's reference genome. A high quality genome assembly of CATD was generated and used to characterize important genes of coconut towards the development of resilient and outstanding varieties especially for added high-value traits. The talk will present the significant results of the project as published in various papers including the first report of whole genome sequence of a dwarf coconut variety. Updates will include the challenges hurdled and specific applications such as gene mining for host insect resistance and screening for least damaged coconuts (thus potentially insect resistant varieties). Genome-wide DNA markers as published and genes related to coconut oil qualitative/quantitative traits will also be presented, including initial molecular/biochemical studies that support nutritional and medicinal claims. A web-based genome database is currently built for ease access and wider utility of these genomics tools. Indeed, a major milestone accomplished by the coconut genomics research team, which was facilitated with the all-out government support and strong collaboration among multidisciplinary experts and partnership with advance research institutes.

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Association between Initial Chest CT or Clinical Features and Clinical Course in Patients with Coronavirus Disease 2019 Pneumonia

  • Zhe Liu;Chao Jin;Carol C. Wu;Ting Liang;Huifang Zhao;Yan Wang;Zekun Wang;Fen Li;Jie Zhou;Shubo Cai;Lingxia Zeng;Jian Yang
    • Korean Journal of Radiology
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    • 제21권6호
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    • pp.736-745
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    • 2020
  • Objective: To identify the initial chest computed tomography (CT) findings and clinical characteristics associated with the course of coronavirus disease 2019 (COVID-19) pneumonia. Materials and Methods: Baseline CT scans and clinical and laboratory data of 72 patients admitted with COVID-19 pneumonia (39 men, 46.2 ± 15.9 years) were retrospectively analyzed. Baseline CT findings including lobar distribution, presence of ground glass opacities, consolidation, linear opacities, and lung severity score were evaluated. The outcome event was recovery with hospital discharge. The time from symptom onset to discharge or the end of follow-up (for those remained hospitalized) was recorded. Data were censored in events such as death or discharge without recovery. Multivariable Cox proportional hazard regression was used to explore the association between initial CT, clinical or laboratory findings, and discharge with recovery, whereby hazard ratio (HR) values < 1 indicated a lower rate of discharge at four weeks and longer time until discharge. Results: Thirty-two patients recovered and were discharged during the study period with a median length of admission of 16 days (range, 9 to 25 days), while the rest remained hospitalized at the end of this study (median, 17.5 days; range, 4 to 27 days). None died during the study period. After controlling for age, onset time, lesion characteristics, number of lung lobes affected, and bilateral involvement, the lung severity score on baseline CT (> 4 vs. ≤ 4 [reference]: adjusted HR = 0.41 [95% confidence interval, CI = 0.18-0.92], p = 0.031) and initial lymphocyte count (reduced vs. normal or elevated [reference]: adjusted HR = 0.14 [95% CI = 0.03-0.60], p = 0.008) were two significant independent factors that influenced recovery and discharge. Conclusion: Lung severity score > 4 and reduced lymphocyte count at initial evaluation were independently associated with a significantly lower rate of recovery and discharge and extended hospitalization in patients admitted for COVID-19 pneumonia.

Chest wall injury fracture patterns are associated with different mechanisms of injury: a retrospective review study in the United States

  • Jennifer M. Brewer;Owen P. Karsmarski;Jeremy Fridling;T. Russell Hill;Chasen J. Greig;Sarah E. Posillico;Carol McGuiness;Erin McLaughlin;Stephanie C. Montgomery;Manuel Moutinho;Ronald Gross;Evert A. Eriksson;Andrew R. Doben
    • Journal of Trauma and Injury
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    • 제37권1호
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    • pp.48-59
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    • 2024
  • Purpose: Research on rib fracture management has exponentially increased. Predicting fracture patterns based on the mechanism of injury (MOI) and other possible correlations may improve resource allocation and injury prevention strategies. The Chest Injury International Database (CIID) is the largest prospective repository of the operative and nonoperative management of patients with severe chest wall trauma. The purpose of this study was to determine whether the MOI is associated with the resulting rib fracture patterns. We hypothesized that specific MOIs would be associated with distinct rib fracture patterns. Methods: The CIID was queried to analyze fracture patterns based on the MOI. Patients were stratified by MOI: falls, motor vehicle collisions (MVCs), motorcycle collisions (MCCs), automobile-pedestrian collisions, and bicycle collisions. Fracture locations, associated injuries, and patient-specific variables were recorded. Heat maps were created to display the fracture incidence by rib location. Results: The study cohort consisted of 1,121 patients with a median RibScore of 2 (range, 0-3) and 9,353 fractures. The average age was 57±20 years, and 64% of patients were male. By MOI, the number of patients and fractures were as follows: falls (474 patients, 3,360 fractures), MVCs (353 patients, 3,268 fractures), MCCs (165 patients, 1,505 fractures), automobile-pedestrian collisions (70 patients, 713 fractures), and bicycle collisions (59 patients, 507 fractures). The most commonly injured rib was the sixth rib, and the most common fracture location was lateral. Statistically significant differences in the location and patterns of fractures were identified comparing each MOI, except for MCCs versus bicycle collisions. Conclusions: Different mechanisms of injury result in distinct rib fracture patterns. These different patterns should be considered in the workup and management of patients with thoracic injuries. Given these significant differences, future studies should account for both fracture location and the MOI to better define what populations benefit from surgical versus nonoperative management.

2차원 바코드를 이용한 오디오 워터마킹 알고리즘 (A digital Audio Watermarking Algorithm using 2D Barcode)

  • 배경율
    • 지능정보연구
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    • 제17권2호
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    • pp.97-107
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    • 2011
  • 본 논문에서는 2차원 바코드를 이용한 오디오 워터마킹 알고리즘을 제안하였다. 삽입되는 워터마크 정보로는 2차원 바코드인 QR 코드를 변형하여 이용하였다. 2차원 바코드가 1차원 바코드에 비하여 많은 정보를 표현할 수 있고, 코드자체가 에러 보정능력을 내재하고 있는 장점을 이용하여 워터마킹 알고리즘의 견고성을 높였다. 또한 부분적인 워터마크 정보의 손실에 대응하기 위하여 직교코드를 이용하여 삽입대역을 확산했으며, 삽입강도 0.7에서 50dB 이상의 우수한 품질을 확보할 수 있었다.