Abstract
CRISPR activation provides an invaluable tool for experimental biologists to convert correlations into causation by directly observing phenotypic changes upon targeted changes in gene expression. With few exceptions, most diseases are caused by complex polygenic interactions, with multiple genes contributing to define the output of a gene network. As such researchers are increasingly interested in tools that can offer not only control but also the capacity to simultaneously upregulate multiple genes. The adaptation of CRISPR/Cas12a has provided a system especially suited to the tightly coordinated overexpression of multiple targeted genes. Here we describe an approach to test for active targeting crRNAs for dFnCas12a-VPR, before proceeding to generate and validate longer crRNA arrays for multiplexed targeting of genes of interest.
Original language | English |
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Title of host publication | Mammalian Synthetic Systems |
Editors | Francesca Ceroni, Karen Polizzi |
Number of pages | 12 |
Publisher | Humana Press |
Publication date | 2024 |
Pages | 193-204 |
Chapter | 8 |
ISBN (Print) | 978-1-0716-3717-3, 978-1-0716-3720-3 |
ISBN (Electronic) | 978-1-0716-3718-0 |
DOIs | |
Publication status | Published - 2024 |
Series | Methods in Molecular Biology |
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Volume | 2774 |
ISSN | 1064-3745 |
Bibliographical note
Publisher Copyright:© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
Keywords
- Cas12a
- CRISPRa
- crRNA assembly
- Multiplexed activation