Title: Development of a Novel CRISPR/Cas9-based Imaging Approach for Real-time Visualization of Specific Genomic Loci in Living Cells

Abstract:

Despite current scientific and medical advancements, we yet lack the knowledge necessary to develop targeted therapies for many diseases, like cancer, for which the precise mechanisms of action are unknown due to technological limitations. Current DNA labelling techniques require the killing and fixation of cells, and often have high noise-to-signal ratios, non-specificity and off-target effects. To understand the mechanism of action of diseases accurately, we need a technology applicable to viable observable cells with minimal noise, high specificity and no off-target effects. Through our studies we developed such a visualization approach that uses a modified form of the CRISPR/Cas9 system, conventionally used for genetic editing, and targets DNA and nuclear elements with the afore mentioned precisions. Some clinically applied treatments like chemotherapy are non-specific; they affect both healthy and diseased tissues. The technique we developed allows us to better understand the pathogenesis of disease-causing nuclear changes by labelling, targeting, observing and imaging them. Consequently, we could develop disease-precise therapies without affecting surrounding healthy tissues. While this tool was used on cervical cancer cells, it may be applied to any living cell. We hope that this approach leads to advancements in the field of precision medicine and would help overcome challenges associated with the development of targeted therapies. Importantly, this approach is of comparable low cost and high efficiency, rendering it economically favourable.

Biography:

Khadeja Alrefaie has completed her BSc in Biotechnology/Medical Technology at California State University Northridge and obtained her MSc in Biotechnology Research Extensive from the University of Queensland Australia, where she developed a novel CRISPR/Cas9 imaging approach to visualize live cells in real-time. Alrefaie believes that translational thinking should be encouraged, and bridging the gap between laboratory, marketplace and clinic has become a necessity. She is currently pursuing her medical and surgical degree (MB, BCh, BAO) from the Royal College of Surgeons in Ireland - Medical University of Bahrain with the intention of combining her clinical skills with her laboratory skills to lead discoveries in the field of biotechnology and biopharmaceuticals

Title: Role of Molecular Diagnostics in Genetic Disorders

Abstract:

Molecular diagnostics is rapidly emerging field of diagnostics . Detection of genomic variants is generally performed to facilitate detection, diagnosis, prognosis, disease monitoring and monitoring, response to therapy. The fruitful interplay among laboratory medicine, genomics knowledge, and technology in the field of molecular genetics has given rise to Molecular Diagnostics. All these factors contribute to the identification and fine characterization of the genetic basis of inherited diseases which, in turn, is vital for the accurate diagnosis. These results allow physicians to not only assess disease predisposition but also to design and implement accurate diagnostic methods and to individualize therapeutic treatment modalities. The various methods or techniques used in the field of Molecular Diagnostics are Next Generation Sequencing (NGS), Sanger Sequencing, Realtime PCR, FISH etc. NGS comprises of whole-genome sequencing (WGS), whole-exome sequencing (WES) and custom designed targeted sequencing. This talk would provide an overview of the molecular diagnostics field and of the molecular diagnostic methods that are being used for patients with genetic disorders. Discussion of these tests and cases will be the highlight of the talk.

Biography:

Ms. Smita Kulkarni Umarji has completed her Masters in Biotechnology from Gulbarga University, Karnataka, India. She is currently working as Senior Technical Executive in Kokilaben Dhirubhai Ambani Hospital, in Genetics department. She has a total of 15 years of experience in the field Molecular Diagnostics. Her core area of competency is Molecular Genetics constituting of oncology and constitutional genetics. She has worked in CAP and NABL accredited laboratories more than a decade which assures her quality of work. She is NABL (National accreditation board) authorized signatory. She also has a certification of CAP for Inspection Team Member.

Title: S-FXS ( B-MS) Study of fragile X Syndrome ( Bell-Martin Syndrome )

Abstract:

Genetic diseases are disorders caused by a defect in the DNA sequence, these disorders are inherited because they are in human sexual cells, sexual cells are responsible for transmitting genetic traits and information from parents to children. Genetic disorders may also sometimes occur due to a DNA sequence defect in physical cells. Genetic diseases: single gene disorders, chromosome disorders

Fragile x syndrome it's also called martin-bell syndrome, it is X- linked dominant inheritance. It is caused by mutation on fmr1 gene that produced protein gene (fmrp), this disorder effect on chromosome X on the long arm at 27.3 region. It is more common in males than females, because it is linked to X chromosome, and we can describe this syndrome by two ways, premutation with 55-200 duplication in triple codon CGG on 5` band, or full mutation with more than 200 duplication in triple codon CGG. until now there is no specific treatment, however there might be a treatment by using the physical and behavioral symptoms or by genetic therapy, these ways are for therapy that can improve patient health. This condition causes several problems, including learning difficulties and cognitive impairment, not only to the central nervous system but also to other physiological imbalances. Distinctive physical qualities are more pronounced as they age, including long face, large ears, prominent jaw and flat feet. In males, the testicles swell after puberty. In addition, not everyone with the syndrome has the same symptoms. The syndrome accounts for half of the mental retardation associated with X-chromosome and is the second most common cause of mental disability after Down syndrome. The first case was discovered in 1938

Meaning of :

Fmrp = fragile mental retardation protein
fmr1= fragile mental retardation protein

Biography:

Haya  has completed her bachelor at the age of 22 years from princess Nourah bint Abdulrahman University. She an intern as a research assistant in king Saud university , college of science , department of plant and microbiology. In addition she is a researcher in king Abdulaziz city for science and technology at life science & environment research institute – National center of agricultural technology

Title: Novel Studies On The Efficacy Of Synthetic Fungicides Against Fusarium Wilt Of Tomato Under Tunnel Farming Condition

Abstract:

Tomato (Lycopersicon  esculentum  Mill.)  is the most  important solanaceous vegetable crop and is one of the most vital  vegetables  economically  in  the  world including Pakistan. It is susceptible to various diseases in tunnel farming conditions which bring huge losses to its production, among them; fusarium wilt is the most destructive disease. This study was conducted to test the efficacy of synthetic fungicides viz.  Metalaxyl+Mancozeb, Copper oxychloride,  Benalaxyl+Mancozeb, Carbendazim and Mancozeb at different concentrations (2, 2.5, 3, 3.5, 4 g/litre water) through soil drench method against fusarium wilt of tomato caused  by  Fusarium  oxysporum  f.  sp.  lycopersici  and also  to  observe  the  impacts  of fungicides on plant height and yield under the tunnel farming condition. The result revealed that Copper oxychloride was significantly effective in all its doses to control the fusarium wilt of tomato as compared to control treatments, the most effective dose was 3 g/l where the disease severity was recorded 6.2% only, followed by Metalaxyl+Mancozeb (4g/l) in which  the  disease  severity was  recorded  9.6% as compared to control (76.6%),  other  fungicides  also  showed  good  results  but Mancozeb alone was not effective,  however it had synergistic effect and could be used as basis with the other  product  to  control  the  fusarium wilt.  Two of the fungicides proved to be the less effective fungicides having no significant effects on plant height and overall yield. The highest plant height was recorded 10.96 and  9.38 as compared to control 3.00  feet,  whereas  the  highest  yield  per  plant  was  recorded  3.97  and  3.67  kg  in  case  of  Copper oxychloride and Metalaxyl+Mancozeb respectively as compared to control 1.60 kg. It is concluded that Copper oxychloride was the most effective fungicide against the fusarium wilt disease among the five tested fungicides under tunnel farming conditions, so should be suggested to be used against this devastating disease.

Biography:

Adnan baloch is a plant pathologist at directorate of vegetable seed productionagriculture research institute, quetta, balochistan. He is also pursued m.sc (agri.) Hons plant pathology from sindh agriculture university, tandojam, pakistan. He conducted several awareness trainings for local farming communities regarding the control of different plant diseases. His research interests are plant pathology, microbiology and plant molecular diseases (fungi, bacteria, virus, nematodes).

Title: Expression study of recombinant p1 protein of local foot and mouth disease virus in yeast cells

Abstract:

Foot and mouth disease is one of the most lethal viral animal diseases worldwide in cleft hoofed animals such as cattle’s, sheep, water buffalo etc. The virus belongs to family Picornaviridae and genus Apthovirus. There are seven serotypes (A, O, C Asia1, and SAT 1-3) of FMDV exist. The serotype O is the most dominant disease in Pakistan. The foot and mouth disease virus protein shell comprises the P1 polyprotein. The P1 polyprotein cleaves into four virus structural proteins VP4, VP3, VP2, and VP1. Among these the VP4 is entirely internal. The capsid protein P1 is associated with the antigenic properties and interact with cell surface receptors.

Inactivated virus vaccine for the prevention of FMD has been commercialized. Vaccination is an only way to overcome this virus. Pichia pastoris is a methylotrophic yeast and possess a strong inducible promoter AOX1.it is a very cost effective eukaryotic expression system with the benefit of post translational modification which is necessary for the proper confirmation of the protein. This study aims to clone the P1 polyprotein gene of local FMDV serotype O strain in methylotrophic yeast Pichia pastoris expression system.

For this purpose P1 gene was modified in Pichia pastoris and copy number was increased in E. coli. The synthetic gene was expressed in P. pastoris GS115 expression system to produce protein. Protein expression was analyzed by using SDS-PAGE and Western Blot. Protein was quantified by using Bradford assay. Indirect ELISA was done to check the expression of the P1 polyprotein. FMDV serotype ‘O’ builted 96 well plates after saturation were used for trapping the antibody. This study would be helpful in construction of a recombinant expression system containing P1 gene from local FMDV (serotype O) in a eukaryotic P. pastoris expression system which will be further used to study the expression of viral protein and to produce recombinant P1 polyprotein as a candidate antigen for the production of recombinant vaccine against FMDV  

Biography:

Muhammad  Asif  Muneer  from University of Veterinary and Animal Sciences Lahore, Pakistan. With full command on High pressure liquid chromatography, solid phase extraction assembly , spectrophotometer, thin layer chromatography and PCR.