Genetic engineering of Mycobacterium leprae to Glow and Grow

麻风分枝杆菌发光和生长的基因工程

基本信息

批准号：
10526890
负责人：
WILLIAM Robert JACOBS
金额：
$ 25.2万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-20 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10526890
关键词：
ATP-Binding Cassette Transporters Achievement Acid Fast Bacillae Staining Method Anabolism Animal Model Bacillus Bacteria Bacteriophages Bioluminescence Carbon Cell Wall Cells Communities Complement Consumption Cosmids DNA DNA Sequence Data Detection Developing Countries Development Diagnosis Disease Electroporation Energy Transfer Engineering Enzymes Evolution Family Dasypodidae Fatty Acids Fireflies Generations Genes Genetic Genetic Engineering Genome Genomics Glucose Goals Green Fluorescent Proteins Growth Harvest Homologous Gene Homoserine Human Immunologics In Vitro Infection Intravenous Iron Lead Leprosy Libraries Light Lipids Luciferases Metabolic Methionine Methods Modeling Mus Mycobacterium leprae Mycobacterium tuberculosis NADH Nude Mice Organ Organism Oxidoreductase Pantothenic Acid Pathway interactions Persons Pharmaceutical Preparations Pharmacotherapy Plasmids Predisposition Process Proteins Pseudogenes Radioactivity Recording of previous events Regimen Regulatory Pathway Reporter Reporting Research Research Personnel Research Technics Siderophores Signal Transduction Source Stains Stigmatization System Techniques Testing Therapeutic Agents Time Tissue imaging Tissues Work acid fast bacteria base cost drug development drug efficacy drug testing foot human disease improved in vivo imaging system intracellular parasitism mycobacterial mycobactins nanoluciferase novel novel therapeutics oxidation pathogen promoter tool uptake vaccine development

项目摘要

Project Summary/ Abstract Leprosy has plagued mankind throughout human history and still afflicts 2 to 3 million people in Developing Countries. Although the leprosy bacillus was the first bacterium reported to be associated with human disease, it has yet never been cultivated on artificial media. It can be grown in the footpads of nude mice or systemically in nine-banded armadillos. With a generation time of 14 days, testing drug regimens are performed by counting acid-fast bacilli which is laborious and time-consuming. The comparison of the M. leprae DNA sequence to M. tuberculosis suggests the inability to culture M. leprae on artificial media might result from the inability to synthesize methionine, pantothenate and mycobactin. Using genetic tools, we have developed for M. tuberculosis, we hypothesize M. leprae can be stably transformed with phage- based integration proficient plasmids. We plan to incorporate a Bioluminescent Resonance Energy Transfer (BRET)-Nanoluc and Near infra-red proteins into the plasmid to detect transformants. Furthermore, we hypothesize that the transformation of M. leprae with the missing methionine and pantothenate biosynthetic genes and the mycobactin transporters will enable M. leprae to grow on artificial media. An M. leprae strain that glows would enhance drug development and the ability to assess the immunological mechanisms that control M. leprae infections. The genetic engineering of M. leprae to grow on artificial media would enhance our understanding of evolutionary processes that lead to obligate intracellular parasitism.

项目摘要/摘要麻风病在整个人类历史上困扰着人类，但仍遭受2至300万人的困扰发展中国家。尽管麻风病芽孢杆菌是据报道的第一个细菌与人类疾病相关，它尚未在人工媒体上种植。可以在裸鼠的脚踏板上生长，或系统地在九个带状的腋窝中生长。有一代 14天的时间，通过计算酸性杆菌进行测试药物方案费力且耗时。 Leprae DNA序列与M.的比较结核病表明，在人工培养基上无力培养麻雀可能是由无法合成蛋氨酸，泛硫酸盐和霉菌素。使用遗传工具，我们有为结核分枝杆菌开发，我们假设Leprae可以通过噬菌体稳定转化。基于熟练质粒的整合。我们计划结合生物发光的共振能量转移（BRET）-NANOLUC和接近红外蛋白进入质粒，以检测转化体。此外，我们假设Leprae M. 蛋氨酸和泛素生物合成基因和菌根转运蛋白将启用M。麻风在人工媒体上生长。发光的M. Leprae菌株会增强药物开发和评估控制M. Leprae的免疫机制的能力感染。 Leprae M.在人工媒体上生长的基因工程将增强我们的了解导致义务细胞内寄生虫的进化过程。