PLASMID CURING ON CANCEROUS DRUG TREATMENT

“The potential for synthetic biology and biotechnology is vast; we all have an opportunity to create the future
together”
~ RYAN BETHENCOURT

Plasmid curing is a laboratory technique used to remove plasmids

from bacterial cells. Plasmids are small, circular DNA molecules
found in bacteria, which often carry extra genes that provide
advantageous traits, such as antibiotic resistance. Plasmid curing
involves treating the bacterial cells with specific agents or
environmental conditions that selectively eliminate the plasmids,
leaving the bacteria with only their chromosomal DNA.

Cancer cells are cells that divide

continually, forming solid tumours or
flooding the blood or lymph with
abnormal cells. Cell division is a normal
process used by the body for growth and
repair. A parent cell divides to form two
daughter cells, and these daughter cells
are used to build new tissue or to replace
cells that have died because of aging or
damage.

Plasmid designed for cancer therapy-: Plasmids used for cancer gene therapy or DNA
vaccination must contain at least one expression cassette that directs the expression of
a protein that will induce the therapeutic effect. After DNA uptake by the cell, it needs
to reach the nucleus, where the gene will direct the therapeutic protein expression in
the same way the cell produces its own proteins. For therapy to be effective, the correct
design and optimization of the plasmid are crucial. For example, if more than one gene
of interest needs to be expressed using a single plasmid, we can even express them
independently (each gene with its own promoter), in a multicistronic system (two or
more genes under the control of the same promoter), or as a fusion protein (a linker
sequence between both sequences may be added). For the multicistronic system, an
internal ribosome entry site (IRES) or a virus-derived biomolecule sequence must be
placed between the different genes.

Usage of protein-: A novel strategy involves

using plasmids that encode monoclonal
antibodies to block different signalling cascades,
such as immune checkpoints or other molecules
expressed on the cell surface or secreted in the
tumour microenvironment.
Genetic Map~

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The chassis organism will be E.coli.
The process of gene transfer can
be summarised in four key steps:

Isolation of gene and Delivery Methods for Plasmids in Cancer

vector (by PCR) Therapeutics~
The plasmids used for gene therapy are usually
Digestion of gene and administered directly into the tumour site to target
vector (by cancer cells. Furthermore, plasmids used for
restriction endonuclease) vaccination are usually administered by mucosal
delivery (where the presence of APCs improves
vaccination efficiency) or by intramuscular,
Ligation of gene and
intradermal, or intratumoural injections to target
vector (by DNA ligase)
either somatic cells, cancer cells, or immune cells
for antigen production.
Selection and expression
of transgenic construct

The simplest form of administration of

plasmids is the injection of naked DNA.
Due to their electrostatic characteristics,
such as their negative charge and size,
plasmids are often administered with
other delivery methods to improve cell
entry. This section reviews some of the
most commonly used delivery methods for
plasmid DNA-based cancer therapies
analysed in in vivo experiments.
Side Effects of Plasmid Curing~

When plasmid DNA-based strategies are translated to clinical trials, different results
have been reported ranging from non-significant anti-tumor responses to effective
therapeutic effects with the induction of antigen-specific cells and tumoural
regression. One of the main challenges in DNA vaccination is the induction of a potent
immune stimulation. Different strategies have been employed to overcome this, such
as consecutive vaccine strategies (known as prime-boost immunization) or the
administration of DNA vaccination in combination with ICB therapy, other
monoclonal antibodies, immunostimulatory molecules, adjuvants, or drugs, among
others.

BY- ARITRIKA MANDAL (XII-A)

NIPOBITHI BISWAS (XII-A)