BIOE 202 Transfection Flashcards

Vocabulary-style flashcards covering the biological, physical, and chemical methods of gene delivery and the specific protocols for mammalian cell transfection from the BIOE 202 lab course.

Transduction

The transfer of genetic material from one organism or microorganism to another by a viral agent.

Transfection

The process of artificially introducing nucleic acids ($DNA$ or $RNA$) or proteins into cells, utilizing means other than viral infection.

Transformation

The transfer of plasmid $DNA$ into a bacterial host to replicate the $DNA$ and verify that the construct is correct.

Transient overexpression

The delivery of $DNA$ to overexpress a protein temporarily for manufacturing or functional studies.

Stable insertion

The delivery of $DNA$ to insert genes into the genome for the purpose of making new cell lines.

Gene silencing

The delivery of agents to suppress the expression of genes for functional studies.

Viral Delivery Methods

Methods utilizing adapted viruses such as Adenovirus, $AAV$, Retrovirus, Lentivirus, and Vaccinia virus for gene delivery.

Physical Methods (Non-Viral)

Methods for gene delivery including gene gun, needle injection, laser irradiation, electroporation, and sonoporation.

Chemical Methods (Non-Viral)

Methods using organic materials (Lipid $NP$, Polymeric $NP$) or inorganic materials ($AuNP$, $CNT$, $MSN$, $MNP$, $QD$) for gene delivery.

Cationic nanoparticle

A structure formed when $DNA$ is mixed with a positively charged amphiphilic substance, resulting in encapsulation.

Liposomes

Positively charged structures that can bind to the negatively charged cell membrane to be endocytosed, eventually releasing $DNA$ into the cell.

Lipofectamine 2000

A high-charge cationic lipid that readily binds anionic substrates; it is characterized as technically simple, highly reproducible, and having low toxicity.

Promoter

A sequence of $DNA$ to which proteins involved in transcription bind to initiate transcription of an $RNA$ transcript from the $DNA$ downstream.

CMV, CAG, and EFS

Three specific synthetic promoter sequences used to drive gene expression in mammalian cells such as $HCT116$.

Green Fluorescent Protein (GFP)

A protein that fluoresces when excited by a specific wavelength of light, serving as an indicator to quantify transfection.

C1V1 = C2V2

The formula used to calculate the required volume of a cell suspension to achieve a target cell count for plating (e.g., transfecting $500,000$ cells in a $24$-well plate).