here we go
Running head: RESEARCH PAPER
Planarians or members of the Phylum Platyhelminthes are well-known for their extreme
survivability and innate regenerative capacity (Alvarado, 2005). They can be exposed
continuously to various pathogenic strains and environmental stresses, and yet a significant
portion of these organisms continue to thrive. Although this amazing survivability could be
attributed to the numerous adaptive mechanisms present in a single organism, one that likely
contributes is the organism’s complement system—an integral part of any living organism’s
innate immune system (Bayer, 2015). It appears that in general, the organism’s immune system
should be intact in order for the regenerative processes to take place.
The current study aims to determine the effects of maiming or hindering some of the
functions of the complement system in Planarians, by interfering with the expression of the A2M
protein. Since the immune system is presumed to have an important role in allowing the
regenerative capabilities of the Planaria to take place, our working hypothesis is as follows:
Interfering with the complement system of Planarians by hindering the expression of the
A2M gene and protein would restrict and even incapacitate the regenerative capabilities of the
said organisms(Ghigo, 2014).
Our negative hypothesis, on the other hand, could be stated as follows:
Interfering with the complement system of Planarians, by hindering the expression of the A2M
gene and protein would have no effect on the regenerative capabilities of the said organisms.
Planarians are integral to the investigation of various biological research problems.
Planarians are primarily used as a model for elucidation of the regenerative ability. The ability to
regenerate and exhibit the extraordinary tissue plasticity comes from stem cells called neoblasts.
Accordingly, “neoblasts replace cells lost to normal physiological turnover while giving rise in
amputated animals to the regeneration blastema, the structure in which missing tissues are
regenerated” (Alvarado, 2012). In other words, these neoblasts that are found in intact
planarians, are the reason for their extraordinary ability to regenerate (Exploratorium, 2016).
However, the purpose of this project is to examine the planarians role of an innate
immune system in the response to pathogens by inhibiting the expression of A2M: the gene was
cloned and used to perform RNA interference experiments (Brandtzaeg, 2016). The RNA
interference technique is widely used in planarian research, which allows for the reduction in the
expression levels of the gene and subsequent signaling pathways that may assume a vital role in
the innate immune function (Gao, 2017). Any change in the worm’s phenotype has been tested
for the ability to display an immune response.
The current study sought to examine the effect of silencing the A2M gene, which may be
part of an innate immune system in the planarian Schmidtea mediterranea (Knakievicz, 2014).
The study had a multistep strategy for the examination of the role of A2M. First, the DNA
sequence of the A2M gene was found in the planarian database using sequence information from
the mouse genome. Then, this RNA was isolated, and cDNA was synthesized. After that, primers
that border the A2M gene were synthesized, and the PCR reaction was done using the cDNA in
order to amplify the A2M gene. The amplified DNA was inserted into a vector with T7
promotors on either side of the inserted gene, and the vector was used to transform E. coli. The
transformed bacteria were used to purify the plasmid, and RNA was synthesized using T7
polymerase. The synthesized dsRNA was puri...