Degree Type

Dissertation

Date of Award

2018

Degree Name

Doctor of Philosophy

Department

Genetics, Development and Cell Biology

Major

Genetics and Genomics

First Advisor

Bing Yang

Abstract

Engineered TALENs (Transcription activator-like effector nucleases) and CRISPR/Cas (clustered regularly interspaced short palindromic repeats/ CRISPR-associated proteins) as genetic toolbox have been widely used in basic and applied biological researches. Here we use TALENs and CRISPR/Cas9 to target miRNA genes and Argonaute (AGO) genes for gene knockouts and study their functions in rice. We successfully knocked out 5 miRNA genes and recovered phenotypic mutant plants using TALENs. The efficient assembling of TALENs constructs and their versatility provided us a very useful toolbox to study miRNA gene functions. In addition, we used CRISPR/Cas9 system to target AGO genes in rice. We successfully generated single-gene knockouts for all 19 AGO genes and several multi-gene mutants of gene clades through multiplex targeting of CRISPR system in rice. Some mutants were characterized at molecular and morphological levels. The results demonstrate TALEN and CRISPR/Cas9 mediated gene editing are robust genetic tools that enable functional genomics.

Copyright Owner

Honghao Bi

Language

en

File Format

application/pdf

File Size

125 pages

Included in

Genetics Commons

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