Chapter title |
Plant Gene Regulation Using Multiplex CRISPR-dCas9 Artificial Transcription Factors
|
---|---|
Chapter number | 12 |
Book title |
Maize
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7315-6_12 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7314-9, 978-1-4939-7315-6
|
Authors |
Levi G. Lowder, Aimee Malzahn, Yiping Qi |
Abstract |
Besides genome editing, the CRISPR-Cas9-based platform provides a new way of engineering artificial transcription factors (ATFs). Multiplex of guide RNA (gRNA) expression cassettes holds a great promise for many useful applications of CRISPR-Cas9. In this chapter, we provide a detailed protocol for building advanced multiplexed CRISPR-dCas9-Activator/repressor T-DNA vectors for carrying out transcriptional activation or repression experiments in plants. We specifically describe the assembly of multiplex T-DNA vectors that can express multiple gRNAs to activate a silenced gene, or to repress two independent miRNA genes simultaneously in Arabidopsis. We then describe a "higher-order" vector assembly method for increased multiplexing capacity. This higher-order assembly method in principle allows swift stacking of gRNAs cassettes that are only limited by the loading capacity of a cloning or expression vector. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 5 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 5 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 47 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 9 | 19% |
Student > Ph. D. Student | 5 | 11% |
Student > Master | 4 | 9% |
Student > Postgraduate | 3 | 6% |
Other | 3 | 6% |
Other | 9 | 19% |
Unknown | 14 | 30% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 18 | 38% |
Biochemistry, Genetics and Molecular Biology | 10 | 21% |
Arts and Humanities | 1 | 2% |
Social Sciences | 1 | 2% |
Chemistry | 1 | 2% |
Other | 1 | 2% |
Unknown | 15 | 32% |