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INTRODUCTION TO GENETIC ENGINEERING | BSM3202��ASSOC. PROF. DR. SITI SARAH OTHMAN

AN INTRODUCTION

WEEK I

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A person whose work is teaching others, or one who is an authority on methods or theories of teaching

EDUCATOR

LECTURERS

Someone who teaches at a college or university

FACILITATOR

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SELECTED PUBLICATIONS:�Hanis Faudzi,, Suet Lin Chia, Raha Abdul Rahim and Sarah Othman. Adherence and internalisation of Lactococcus lactis M4 towards human colorectal cancer cell line, Caco-2. 2021, Malaysian Journal of Microbiology. Vol 17(3).�M. A. Farahani, M. Zamri-Saad, A. R. Raha, P. Chumnanpuen, T. Ekobon, S. Othman. Molecular Characterisation of the GdhADerivative of P. multocida B:2. 2021, Pertanika J. Trop. Agric. Sci. 44 (1), 171 - 192Cha Yee Kuen, Tieo Galen, Sharida Fakurazi, Siti Sarah Othman, Mas Jaffri Masarudin . (2020). Increased Cytotoxic Efficacy of Protocatechuic Acid in A549 Human Lung Cancer Delivered via Hydrophobically Modified-Chitosan Nanoparticles As an Anticancer Modality. Polymers.(12) 9:1951.Theeraphol Jatuponwiphat, Pramote Chumnanpuen, Sarah Othman, E Teerasak, Wanwipa Vongsangnak. (2019). Iron-associated protein interaction networks reveal the key functional modules related to survival and virulence of Pasteurella multocida. Microbial Pathogenesis. 127 (257-266).�Habibah Faroque, Yi Siang Lau, Chee Xhian Yong, Raha Abdul Rahim, Suet Lin Chia, and Sarah Othman (2018). Bactofection of SW620 cell by Lactococcus lactis M4. Asia Pacific Journal of Molecular Biology and Biotechnology. (26) 1:29-41.�Hanis Faudzi, Chia Suet Lin, Raha Abdul Rahim, Sarah Othman (2018). Lactococcus lactis: LAB model organism for bacteria-mediated therapeutic strategies. Asia Pacific Journal of Molecular Biology and Biotechnology. (26) 1: 1-10.�Sengkar Yap, Zunita Zakaria, Siti Sarah Othman, Abdul Rahman Omar, (2017). In vitro treatment of lipopolysaccharide increases the invasion of Pasteurella multocida serotype B:2 into bovine aortic endothelial cell. Journal of Veterinary Science.�NM Kamal, M Zamri-Saad, MJ Masarudin, S Othman, (2017). Interaction between Pasteurella multocida B: 2 and its derivatives with bovine aortic endothelial cell (BAEC). BMC Veterinary Research 13 (1), 186.�AA Bitrus, Z Zunita, SK Bejo, S Othman, NAA Nadzir, (2017). In vitro transfer of methicillin resistance determinants mec A from methicillin resistant Staphylococcus aureus (MRSA) to methicillin susceptible Staphylococcus aureus (MSSA). BMC Microbiology 17 (1), 83.�Othman S, Roe AJ, Parton R, Coote JG (2013) Use of a Dual Reporter Plasmid to Demonstrate Bactofection with an Attenuated AroA-Derivative of Pasteurella multocida B:2. PLoS ONE 8(8): e71524. �Othman S., R. Parton, and J. Coote. (2012). Interaction between mammalian cells and Pasteurella multocida B:2. Adherence, invasion and intracellular survival. Microbial pathogenesis 52, 353-358. �

ASSOC. PROF. DR. SITI SARAH OTHMAN

PhD. (Glasgow), Infection & Immunity

sarahothman@upm.edu.my

CURRENT RESEARCH:

My research focuses on the utilization of ruminant pathogens and probiotic bacteria for DNA vaccine delivery for veterinary diseases or for cancer immunotherapy. Current research endeavors include investigating the mechanism of host-pathogen interactions between ruminant pathogen and host’s cells, development of multi-valent vaccine for veterinary diseases and colorectal cancer immunotherapy via bactofection.

RESEARCH INTERESTS:

  • Veterinary Vaccine delivery
  • Host-pathogen Interactions
  • Cancer Immunotherapy

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SELECTED PUBLICATIONS:�Yusof Nurliyana Mohd, Mohamed Noor Fuadi Natasha Nur Afiqah, Osman Mohd Azuraidi, Mohd Lila Mohd Azmi, Abd Rahman Nik Mohd Afizan Nik . (2020). Viruses, the vehicle for cancer gene therapy: A Review. African Science Publications.�MAF Yahaya, MAM Lila, S Ismail, M Zainol, NAR Afizan. 2019. Tumour-associated macrophages (TAMs) in colon cancer and how to re-educate them. Journal of immunology research.Ruzila Ismail, Zeenathul Nazariah Allaudin Rasedee Abdullah, Mohd-Azmi Mohd Lila, Sheikh-Omar Abdul Rahman, Nik-Mohd-Afizan Nik Abd. Rahman. (2016) Combination of VP3 and CD147-knockdown enhance apoptosis and tumor growth delay index in colorectal tumor allograft. BMC cancer 16 (1).�NAR Nik-Mohd-Afizan, Y Watanabe, N Higo, I Takashima. (2015) Brain Stimulation 8 (2). Effect of the Transcranial Direct Current Stimulation on Permeability of Blood Brain Barrier in the Rat. �Firdaus Romli, Noorjahan Banu Alitheen, Muhajir Hamid, Ruzila Ismail, Nik Abd Rahman (2013) Current techniques in reprogramming cell potency. Journal of cellular biochemistry114 (6) 1230-1237

ASSOC. PROF. DR. NIK MOHD AFIZAN NIK ABD RAHMAN

Ph.D. (UPM), Cancer Biology, Vaccine Technology

m.afizan@upm.edu.my

CURRENT RESEARCH:

Our research focuses on the development of cancer vaccines through gene therapy and cell reprogramming approaches to delay and stop cancer cells growth. Recombinant viral vectors harbouring therapeutic genes known as gene therapy used in tackling tumor progression. Cells reprogramming and blood barrier present a novel approach for understanding disease progression and drug effectiveness.

RESEARCH INTERESTS:

  • Cancer Therapeutic Vaccine
  • Cellular Reprogramming
  • Blood Barrier

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Are you inspired?�Can you inspire? ☺

Perhaps..you’re expired? ☹

How do you stay inspired?

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The #5things

  1. 70% to 30% ratio
  2. Curiosity – Driven Education
  3. Projection to derive hypothesis
  4. Ethics – Scientist in Training (SiT)
  5. Enjoy the ride & have FUN!! ^_^

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THE REAL DEAL

  • 1st TEST: 10%
  • 2nd TEST: 10%
  • SCL: 40%
  • FINAL: 40%
  • Practical report : 5%

  • PBL : 15%

  • Laboratory exam : 10%

  • Quiz & Tutorial : 10%

Lecture notes, practical manual, quizzes, additional reading material are available through PUTRA BLAST

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WHAT IS RECOMBINANT DNA?

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The Most Popular Sheep in the World!!

Facts about Dolly:

  • 1st mammal cloned from the udder cell of an adult sheep — after 276 attempts
  • Dolly was born in July 1996, however researchers announced Dolly's existence on Feb. 22, 1997
  • Dolly had a total of six lambs. Her first lamb, Bonnie, was born in April 1998.
  • Dolly was put to sleep on 14th February 2003, at the age of six after she was diagnosed with several diseases including arthritis and lung cancer.

Dolly the Sheep with her first born lamb, called Bonnie.The Roslin Institute, The University of Edinburgh

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DEFINITION

Molecules of DNA from two different species that are inserted into a host organism to produce new genetic combinations that are of value to science, medicine, agriculture, and industry.

TECHNIQUES

Recombinant DNA technology will produce clones through the production of many copies of a single fragment of DNA, such as a gene, creating identical copies that constitute a DNA clone.

In practice the procedure is carried out by inserting a DNA fragment into a small DNA molecule and then allowing this molecule to replicate inside a simple living cell such as a bacterium.

(Britannica by Anthony J.F. Griffiths, Professor Emeritus of Botany, University of British Columbia, Vancouver)

Recombinant DNA Technology

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What is Genome Editing?

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“The 2020 selection for the Nobel Prize in Chemistry goes to two scientists who share credit for identifying and developing a revolutionary method of genome editing — one that has allowed researchers to modify and investigate the genomes of microbial, plant and animal cells with an ease, precision and effectiveness that would have been unfathomable even a decade ago.

Emmanuelle Charpentier of the Max Planck Unit for the Science of Pathogens Institute for Infection Biology and Jennifer Doudna of the University of California, Berkeley have been recognized for their work on CRISPR/Cas9 genome editing — a technique routinely called CRISPR (clustered regularly interspaced short palindromic repeats) for short and often referred to as “genetic scissors.” This award marks the first time that two women have been award a Nobel Prize for science.”

~geneticliteracyproject.org

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It’s in your hands!

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  1. The basic techniques for cloning and manipulation of genes
  2. Procedure for cloning, screening and identification of heterologous genes
  3. Cloning vectors
  4. Restriction of vectors and insertion of DNA and process involved in transformation
  5. PCR technology
  6. DNA sequencing technology
  7. Ethical issues in relation to recombinant DNA technology
  8. Practical exercises in order to plan and execute experiments involving recombinant DNA

Course Synopsis

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INTRODUCTION TO GENETIC ENGINEERING

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WHAT DO YOU THINK?

  • Using the annotation in Zoom, please indicate your opinion on the questions given to you in the next slide?
  • You can have as many answers as possible per question. Annotate your answer under the designated question.

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WHAT DO YOU THINK?

  1. What do you understand by the phrase “in vitro gene manipulation”?

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WHAT DO YOU THINK?

2. Which one is the most economical model to be used in gene manipulation; virus, bacteria, yeast, plant or animal? Why?

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  1. Explain the concept and importance of genetic engineering (C2)
  2. Identify how a plasmid contributes to a selective advantage for a bacterial cell (C3)

Learning Outcomes

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Recombinant DNA Technology

The ability to propagate foreign DNA

Gene Manipulation

(GENETIC ENGINEERING)

The ability to cross natural species barriers

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RECOMBINANT DNA – GENE MANIPULATION

WHAT DO WE NEED?

HOST

CAR??

TARGET GENE

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Genetic Manipulations : In vivo Techniques

(C) Requirement for growth compounds

Prototroph: a microorganism that has the same nutritional requirements as the parent organism

Auxotroph:

a mutant strain of microorganism having nutritional requirements additional to those of the normal organism (e.g. proline: cannot synthesize proline, cannot grow without proline in media)

  1. Sensitivity to chemicals

Antibiotics (Ampicillin, chloramphenicol)

(B) Ability/inability to break down carbon compounds

lac- (cannot break down lactose),

gal- (cannot break down galactose)

1. Use of microorganisms

a) easy and simple to grow

b) rapid culture (genetically pure strains )

c) availability of identification and selection techniques

2. Selection of bacterial mutants:

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APPLICATION

GENE MAPPING

GENE EDITING

MICROARRAY

EPIGENETICS

IMPORTANCE OF RECOMBINANT DNA

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Lack of it is believed to be responsible for killing more children than HIV - around 2,000 deaths a day. On a global scale, about a third of children under five suffer from the condition which can also lead to blindness.

Golden Rice has not been made available to those for whom it was intended in the 20 years since it was created because of political and ignorance issues.

Had it been allowed to grow in these nations, millions of lives would not have been lost to malnutrition, and millions of children would not have gone blind. ~ The Guardian, UK ~

Golden Rice has been genetically modified to contain a higher concentration of vitamin A (1999).

Aim is to aid people in the developing countries to counter blindness and other diseases because of vitamin A deficiency.

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Recombinant human insulin has been produced in 1982 predominantly using E. coli and Saccharomyces cerevisiae for therapeutic use in human. 

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GENE THERAPY

GENOME EDITING

DESIGNER BABIES?

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Stress tolerant maize in Africa –AgriOrbit

Drought resistant higher yielding GM rice

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ACTIVITY I�“BE A SCIENCE COMMUNICATOR

  1. Get into a group of 5 (preferably mix genders and races)
  2. Search for an establish application of genetic engineering [it can be product (ex: insulin), application, system or policy]
  3. Understand the scientific concept behind the application
  4. Create a maximum of 1-minute TikTok video about the chosen application in layman term (suitable for public understanding)

Objective: to educate and promote STEM education among the public

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ACTIVITY I�BE A SCIENCE COMMUNICATOR

  • Submit your group details, video title and content details in the online google excel link provided in the PutraBlast.
  • Your video MUST be submitted by Monday the 8th of November 2021 before 10 am.
  • Your video will be available in GE online platform for external and peer judging session.