RECOMBINANT DNA METHODOLOGIES

Course objectives

-General objectives The goal of this course is to provide the students with the knowledge of the recombinant DNA techniques, in order to allow a critical approach and analysis of molecular- experiments.The use of recombinant DNA techniques in basic as well as applied (biomedical and agronomic) research will be described. Both the nucleic acids and proteins basic techniques, up to new ones for "omics" studies will be addressed. Examples will be taken from the current scientific literature and updated every year. The ethical implications and the most recent discoveries in this field will be also discussed. - Specific objectives Knowledge and understanding - Students will get acquainted with the essential concepts and techniques used in genetic engineering Applying knowledge and understanding - Students will acquire the ability to interpret and discuss current research being performed genetic engineering Making judgements Students will learn to critically discuss and evaluate the possible ethical implications of a study in the field of genetic engineering Communication skills –Students enrolled in the course will deliver several short (~10-15 minute) oral presentations on selected scientific papers throughout the course. This will enhance the communication skills of all the students attending this course. Learning skills - Students will acquire not only the scientific basis of genetic engineering, but also the terminology and the methodologies used in this field. This skills will accompany the students during the rest of their academic career.

Channel 1
PAOLA VITTORIOSO Lecturers' profile

Program - Frequency - Exams

Course program
Genetic engineering basic Tecniques: restriction/modification enzymes, linkers, adapters. DNA probes, Southern, Northern, Western blot. Polymerase chain reaction (PCR), Real time and applications. DNA sequencing: manual and automatic sequencing. Next Generation Sequencing (NGS) and applications. Cloning tecniques: traditional cloning, Gateway and Golden technologies. Transformation of pro/eukaryotic cells. Genomic and cDNA lybraries. Screening Strategies. Expression analysis: Northern blot, semi-quantitative and competitive RT-PCR, qRT-PCR, RNA-seq. Microarray and Affymetrix technologies. Reporter genes, promoter analysis. Heterologous expression in E.coli, S.cerevisiae and in eukaryotic cells. DNA-proteins interaction: one-hybrid, south western, Electrophoretic Mobility Shift Assay (EMSA), footprinting, Chromatin Immuno Precipitation (ChIP). ChIP-on-chip. ChIP-seq. RNA-proteins interaction: RNA pull-down, RIP, CLIP. Protein-protein interactions: yeast two, three, four-hybrid, co-immunoprecipitation (co-ip), pull down assays. Bimolecular fluorescence complementation. Fluorescence resonance energy transfer (FRET), Split-ubiquitin. DNA fingerprinting, genetic markers (VNTR, STR, SNP)."Chromosome walking". Animal cells transformation. Animal cloning: from Dolly to Macaque Monkeys by Somatic Cell Nuclear Transfer (SCNT). Plant transformation, plant genetic engineering. Genetic Modified Organisms (GMO). Strategies to study gene function: forward and reverse genetics. RNA interference and applications. Targeting-induced local lesions in genomes (TILLING). Genome editing: from Talen to Crisp-Cas9. Synthetic biology. Biosensors and applications. Optogenetics.
Prerequisites
There are no prerequisites for this course. However, it is suitable that the student has knowledge of molecular biology, genetics and biochemistry. The course does not require mandatory attendance.
Books
Terry A. Brown Biotecnologie Molecolari. Ed. Zanichelli. 2° ed. italiana. Libro multimediale. Zlatanova J. and van Holde K.E. Biologia molecolare (chapt. 5 and methods described in several chapters). Lizabeth A. Allison Fondamenti di biologia molecolare (cap.13, 14), Ed. Zanichelli. 2° ed. italiana. Libro multimediale. Files .pdf available at http://elearning2.uniroma1.it/
Teaching mode
Lessons will take place in class, unless there are restrictions imposed by the current pandemic situation. In any case, it will be possible to connect to the google meet platform for students unable to attend the lesson
Frequency
Although attendance is not mandatory, it is strongly recommended.
Exam mode
The exam will take place orally and will include a discussion of the topics covered in class with the development of a small research project. Furthermore, it will be possible to discuss a research article previously chosen by the teacher.
Lesson mode
Lessons will take place in class, unless there are restrictions imposed by the current pandemic situation. In any case, it will be possible to connect to the google meet platform for students unable to attend the lesson
  • Lesson code1019207
  • Academic year2025/2026
  • CourseBiology
  • CurriculumGenetico-molecolare
  • Year3rd year
  • Semester2nd semester
  • SSDBIO/11
  • CFU6