Date: Tuesday, 23 August 2016, 10 am (Manila Time)
The genome found in every cell of our body contains over 20 thousand genes and over 3 billion letters of DNA that sustains life, shapes who we are and determines our risks of having a disease. CRISPR/Cas (clustered regularly interspaced palindromic repeats) is a recently discovered antiviral defence system in bacteria that has become the favorite set of tools to edit and correct any diseased genome and change any sequence of DNA in precisely chosen genomic location performed not in a test tube but within the nucleus of our living cell. CRISPR/Cas is a group of molecular scissors comprising of proteins complexed with small guide/tracer RNAs that can precisely detect targeted sequence of DNA for binding, cutting or producing single or double-strand breaks (DSBs). DSBs are repaired imprecisely by non-homologous end-joining (NHEJ) to generate random insertion or deletion mutations, or repaired precisely by homology directed repair (HDR) in the presence of a new piece of exogenously-introduced donor template DNA that can pair up with the broken ends to allow insertion, substitution or fix any defective gene. This technology has been used to help researchers better understand the functions of certain genes and produce specific disease models. In cancer for example, researchers can take the patients own immune cells and edit their genes to recognize and attack the cancer cells when put back into the patient’s system. Generating a double-strand break (DSB) is essential to promote exchange of genetic materials. Although DSBs are harmful and considered as the most deleterious among all DNA lesions, they are the driving force behind any genomic perturbation or editing and are required to induce occurrence of DNA repair processes to achieve genome stability. Tips and considerations in utilizing CRISPR/Cas9 to produce precise DNA binding or DNA breaks are discussed focusing on how to improve results to cater towards specific genomic engineering application.
Mark I.R. Petalcorin, PhD
Associate Professor of Biochemistry
PAPRSB – Institute of Health Sciences
Universiti Brunei Darussalam
About the Speaker
Dr. Mark PETALCORIN is currently an Associate Professor of Biochemistry at Universiti Brunei Darussalam, PAPRSB Institute of Health Sciences. He completed his Bachelor’s degree in Agronomy/Plant Breeding at University of the Philippines at Los Baños and his masters/PhD degrees in Biochemistry/Molecular Biology at Osaka University Japan. He worked as a postdoc for 3 years working on Drug metabolizing enzymes at Institute of Pharmaceutical Science, Kings College London and spent another 12 years as a senior scientist at Clare Hall Laboratories, Francis Crick Institute in London, UK investigating DNA repair pathways after DNA double strand breaks. He is an expert in genome editing using model organisms and mammalian cells utilizing CRISPR-Cas system and BACs recombineering via nonhomologous end joining and homologous recombination. His research interests cover cancer proteomics using mass spectrometry and peptide arrays, biochemistry of protein-protein, protein-DNA or protein-substrate interactions and the genetics of model organism C. elegans focusing on genome instability, meiosis, homologous recombination and DNA damage response.
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