RNAi C elegans research poster 2021 | Assignments Biology

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C. elegans is a model organism commonly used in the study of

human diseases and genetics. In this experiment ribonucleic

acid interference (RNAi) will be induced in C. elegans by

feeding to determine embryonic lethality of a gene of choice.

RNAi refers to a cellular process in which different RNA

molecules are utilized to silence selected genes, without altering

the DNA. The gene of choice was F49D11.1, which encodes for a

protein used in mRNA splicing. It is expressed throughout every

embryonic developmental stage, the muscle cells, germ line, and

neurons.'The objective of this study is to investigate the effects

of silencing F49D11.1 then determining if the absence of this

gene leads to'embryonic lethality. After RNAi had concluded

and the hatched progeny were scored, it was found that Day 1

plates containing affected worms had a 74.3% lethality rate,

just 2% under the average for the positive control. Day 2 plates

containing the affected worms had a 100% lethality rate. The

results of our investigation were highly conclusive that silencing

of'F49D11.1 causes embryonic lethality.'

ABSTRACT

DISCUSSION

Caenorhabditis elegans'is a species of small (1mm) free living

nematodes that shareanurbilaterianancestor with humans. As aresult

of geneconservation, the form,function, and genetics of C. elegans

tissues areanalogous to those in humans (2). Thismakes the species

highly relevant for genetics research and the studyofhuman diseases

including cancer, neurodegenerative diseases and developmental

syndromes. The objective of this project was to understand how gene

expression is controlled using RNAi. A simple nervous system,

transparent body,and a three-day life cyclemakeC. elegans an

exceptionalmodel organism forunderstanding just one of many

biologicalfunctions regulated by RNAi (1). RNAi evolved to exist

naturally as adefense against viruses and for the purpose of gene

regulation andcan be used in lab to prevent the production of proteins

encoded bythe F49D11.1 gene. The purpose of this research is to

investigatetheeffects of silencing the F49D11.1 mRNA splicing gene.

INTRODUCTION

•Streaking out RNAi clone: streak out RNAi clones on

agar plates with tetracycline and ampicillin

•Inoculating an Overnight Culture: transfer bacteria into

LB broth then shake and incubate overnight at 37° C

•Plasmid Purification: alkaline lysis procedure with the E.

coli plasmid DNA

•Gel Electrophoresis: run an agarose gel with the plasmid

DNA to determine quality, quantity, and size

•Sequence Analysis: review the DNA sequence file of the

plasmid DNA using Basic Local Alignment Search Tool

(BLAST) to confirm the quality and identity of the DNA

sequence

•RNAi by Feeding: pick 3-4 L4 worms to two agar plates

(Day 1 plate and Day 2 plate) with bacteria that contain a

L4440 plasmid with the F49D11.1 gene to ingest

its’dsRNA which will induce RNAi

•Control Plates: Set up a negative (SMD1) and positive

(Y18D10A.20) control plate

•Scoring the Progeny: count the unhatched eggs and L1

worms then calculate the viability of each plate

•When the expression of a certaingene issuppressed,

the function of that gene can beobserved if thereare

differences in theexperimentalC. elegans plates vs.

the negative control plate

METHODS

RESULTS

As expected,Day 1 negative control displayed a high viability and low lethality. Day 2 positive control was not as lethal as expected. Day 1

F49D11.1 silenced progeny had viability and lethality comparable to the positive control. Out of all plates, Day 2 F49 silenced progeny had

theleast viability with many unhatched eggs and a high lethality rate.

REFERENCES

Since the negative control had a low embryonic lethality and the positive control had a high embryonic lethality, we can conclude that both controls worked as they were supposed to. When the

experimental F49D11.1 gene was knocked down in the worms, there was a high embryonic lethality comparable tothe rates of the positive control. Based on the data, it can be concluded that the F49D11.1

gene had a functional effect since its knockdown resulted in embryonic lethality. The implication of this is that the F49D11.1 gene is essential for embryonic viability and development. Somelimitations

ofthis experimentincludedtime constraints due to limited lab timeas well as deviation between the researchers' accuracy of scoring the progeny.

Faculty Sponsor: Jacqueline Gapinski (Ph.D.) | Bellevue College, Department of Molecular Biosciences

Sorrel Robertson, Samantha Tian, & Jaime Mohammed

Effects of F49D11.1 Gene Silencing in C. elegans

Figure 2. Graphed embryonic viability and lethality of negative, positive, and F49 RNAi.

Figure 1. Day 2 plates with F49D11.1 worms (Plates A & B)

1. Jnimon. (2013, November 13). Model organisms: Shining examples for simple,

effective biology research. NASA. Retrieved December 10, 2021, from

https://blogs.nasa.gov/ISS_Science_Blog/2013/11/13/model-organisms-shining-examp

les-for-simple-effective-biology-research/

.

2. Lundquist, E. A. (n.d.). What is C. elegans and why do scientists use it to study

human development and disease? Why study C. elegans? Retrieved December 10,

2021, from

http://www.people.ku.edu/~erikl/Lundquist_Lab/Why_study_C._elegans.html.

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RESEARCH POSTER PRESENTATION DESIGN © 2015 www.PosterPresentations.com C. elegans is a model organism commonly used in the study of human diseases and genetics. In this experiment ribonucleic acid interference (RNAi) will be induced in C. elegans by feeding to determine embryonic lethality of a gene of choice. RNAi refers to a cellular process in which different RNA molecules are utilized to silence selected genes, without altering the DNA. The gene of choice was F49D11.1, which encodes for a protein used in mRNA splicing. It is expressed throughout every embryonic developmental stage, the muscle cells, germ line, and neurons. The objective of this study is to investigate the effects of silencing F49D11.1 then determining if the absence of this gene leads to embryonic lethality. After RNAi had concluded and the hatched progeny were scored, it was found that Day 1 plates containing affected worms had a 74.3% lethality rate, just 2% under the average for the positive control. Day 2 plates containing the affected worms had a 100% lethality rate. The results of our investigation were highly conclusive that silencing of F49D11.1 causes embryonic lethality.

ABSTRACT

DISCUSSION

Caenorhabditis elegans is a species of small (1mm) free living nematodes that share an urbilaterian ancestor with humans. As a result of gene conservation, the form, function, and genetics of C. elegans tissues are analogous to those in humans (2). This makes the species highly relevant for genetics research and the study of human diseases including cancer, neurodegenerative diseases and developmental syndromes. The objective of this project was to understand how gene expression is controlled using RNAi. A simple nervous system, transparent body, and a three-day life cycle make C. elegans an exceptional model organism for understanding just one of many biological functions regulated by RNAi (1). RNAi evolved to exist naturally as a defense against viruses and for the purpose of gene regulation and can be used in lab to prevent the production of proteins encoded by the F49D11.1 gene. The purpose of this research is to investigate the effects of silencing the F49D11.1 mRNA splicing gene.

INTRODUCTION

(^) Streaking out RNAi clone: streak out RNAi clones on agar plates with tetracycline and ampicillin
(^) Inoculating an Overnight Culture: transfer bacteria into LB broth then shake and incubate overnight at 37° C
(^) Plasmid Purification: alkaline lysis procedure with the E. coli plasmid DNA
(^) Gel Electrophoresis: run an agarose gel with the plasmid DNA to determine quality, quantity, and size
(^) Sequence Analysis: review the DNA sequence file of the plasmid DNA using Basic Local Alignment Search Tool (BLAST) to confirm the quality and identity of the DNA sequence
(^) RNAi by Feeding: pick 3-4 L4 worms to two agar plates (Day 1 plate and Day 2 plate) with bacteria that contain a L4440 plasmid with the F49D11.1 gene to ingest its’ dsRNA which will induce RNAi
(^) Control Plates: Set up a negative (SMD1) and positive (Y18D10A.20) control plate
(^) Scoring the Progeny: count the unhatched eggs and L worms then calculate the viability of each plate - (^) When the expression of a certain gene is suppressed, the function of that gene can be observed if there are differences in the experimental C. elegan s plates vs. the negative control plate

METHODS

RESULTS

As expected, Day 1 negative control displayed a high viability and low lethality. Day 2 positive control was not as lethal as expected. Day 1 F49D11.1 silenced progeny had viability and lethality comparable to the positive control. Out of all plates, Day 2 F49 silenced progeny had the least viability with many unhatched eggs and a high lethality rate.

REFERENCES

Since the negative control had a low embryonic lethality and the positive control had a high embryonic lethality, we can conclude that both controls worked as they were supposed to. When the

experimental F49D11.1 gene was knocked down in the worms, there was a high embryonic lethality comparable to the rates of the positive control. Based on the data, it can be concluded that the F49D11.

gene had a functional effect since its knockdown resulted in embryonic lethality. The implication of this is that the F49D11.1 gene is essential for embryonic viability and development. Some limitations

of this experiment included time constraints due to limited lab time as well as deviation between the researchers' accuracy of scoring the progeny.

Faculty Sponsor: Jacqueline Gapinski (Ph.D.) | Bellevue College, Department of Molecular Biosciences

Sorrel Robertson, Samantha Tian, & Jaime Mohammed

Effects of F49D11.1 Gene Silencing in C. elegans

Figure 2. Graphed embryonic viability and lethality of negative, positive, and F49 RNAi. Figure 1. Day 2 plates with F49D11.1 worms (Plates A & B)

Jnimon. (2013, November 13). Model organisms: Shining examples for simple, effective biology research. NASA. Retrieved December 10, 2021, from https://blogs.nasa.gov/ISS_Science_Blog/2013/11/13/model-organisms-shining-examp les-for-simple-effective-biology-research/ .
Lundquist, E. A. (n.d.). What is C. elegans and why do scientists use it to study human development and disease? Why study C. elegans? Retrieved December 10, 2021, from http://www.people.ku.edu/~erikl/Lundquist_Lab/Why_study_C._elegans.html.

RNAi C elegans research poster 2021, Assignments of Biology

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Partial preview of the text

Download RNAi C elegans research poster 2021 and more Assignments Biology in PDF only on Docsity!

ABSTRACT

DISCUSSION

INTRODUCTION

METHODS

RESULTS

REFERENCES

Since the negative control had a low embryonic lethality and the positive control had a high embryonic lethality, we can conclude that both controls worked as they were supposed to. When the

experimental F49D11.1 gene was knocked down in the worms, there was a high embryonic lethality comparable to the rates of the positive control. Based on the data, it can be concluded that the F49D11.

gene had a functional effect since its knockdown resulted in embryonic lethality. The implication of this is that the F49D11.1 gene is essential for embryonic viability and development. Some limitations

of this experiment included time constraints due to limited lab time as well as deviation between the researchers' accuracy of scoring the progeny.

Faculty Sponsor: Jacqueline Gapinski (Ph.D.) | Bellevue College, Department of Molecular Biosciences

Sorrel Robertson, Samantha Tian, & Jaime Mohammed

Effects of F49D11.1 Gene Silencing in C. elegans