Abstract

Of the entire human genome, 90% of all genetic information is transcribed but only a fraction of that subsequent RNA is translated into proteins. RNAs which are not translated into proteins are deemed non-coding RNAs. Little is known about this large category of noncoding RNAs, although they perform a variety of functions within the cell. RNA crystallography is used to study RNA tertiary structure, which gives insight to the function of these non-coding RNAs. However, complications associated with RNA crystallography arise due to RNA's lack of surface functional group diversity, flexible tertiary structure, and conformational heterogeneity. A novel technique, Chaperone-assisted RNA crystallography (CARC), can greatly improve the success in crystallization of RNA. With this technique, synthetic antibodies called Antigen Binding Fragments (Fabs) are employed as crystallization chaperones to promote the structure elucidation of certain target RNAs. To identify Fabs that complex with RNA molecules of interest, we constructed a randomized library of synthetic antibodies enriched in ligand binding regions with tyrosine, serine, glycine, and arginine residues. This Fab protein library was constructed with a minimal codon design, and then screened against a variety of RNA targets. Several Fabs have been identified and isolated through phage display selection against three RNA targets. These Fabs were expressed and biochemically characterized for their binding affinities and specificities. To date, five Fabs have been identified and they have outstanding capabilities in binding their specific RNA antigen.

Notes

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Thesis Completion

2012

Semester

Spring

Advisor

Ye, Jingdong

Degree

Bachelor of Science (B.S.)

College

Burnett School of Biomedical Sciences

Degree Program

Molecular Biology and Microbiology

Subjects

Dissertations, Academic -- Medicine;Medicine -- Dissertations, Academic

Format

PDF

Identifier

CFH0004167

Language

English

Access Status

Open Access

Length of Campus-only Access

None

Document Type

Honors in the Major Thesis

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