Degree Type

Dissertation

Date of Award

2019

Degree Name

Doctor of Philosophy

Department

Genetics, Development and Cell Biology

Major

Genetics

First Advisor

Bing Yang

Abstract

Genome editing technologies have revolutionized the world of biology. Genome editing for site-specific genomic alterations started with engineered meganucleases, followed by programmable zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat associated protein (CRISPR/Cas). The latter consists of CRISPR-Cas9, -Cpf1(Cas12a), -C2c1(Cas12b), -C2c2(Cas13a) and CRISPR derived base editor (Adenine and Cytosine Base Editor) are state-of-the-art targeted mutagenesis toolkits for basic and applied research. Users have more options to edit, delete, insert, inactivate, activate, repress, label, methylate and acetylate the genetic elements (DNA or RNA) as they need.

In this dissertation, I will present results from some proof-of-concept experiments and demonstrate the genome editing technologies including TALENs, CRISPR/Cas9 and CRISPR/Cas12a that function efficiently in maize, sorghum, and rice. First, the application of custom-tailored TALENs technology to target the glossy2 gene in maize was demonstrated. The methods from assembly of TALEN repeats to analysis and characterization of the inheritability and phenotype of glossy2-edited plants in T1 generation will be described. Next, I will showcase an efficient Agrobacterium tumefaciens mediated CRISPR/Cas9 genome editing method in maize. On this basis, I extrapolated this robust technology to another essential crop, sorghum that feeds and fuels the world using different version of CRISPR/Cas9 system. Finally, I will outline a protocol, showing the application of an optimized CRISPR/Cas12a technology in rice.

In a nutshell, genome-editing tools benefit us with the knowledge of basic biology. By leveraging genome-editing technology, high quality and quantity crops can be produced by farmers in the near future.

Copyright Owner

Si Nian Char

Language

en

File Format

application/pdf

File Size

173 pages

Included in

Genetics Commons

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