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- Title
PARA: A New Platform for the Rapid Assembly of gRNA Arrays for Multiplexed CRISPR Technologies.
- Authors
Yuan, Guoliang; Martin, Stanton; Hassan, Md Mahmudul; Tuskan, Gerald A.; Yang, Xiaohan
- Abstract
Multiplexed CRISPR technologies have great potential for pathway engineering and genome editing. However, their applications are constrained by complex, laborious and time-consuming cloning steps. In this research, we developed a novel method, PARA, which allows for the one-step assembly of multiple guide RNAs (gRNAs) into a CRISPR vector with up to 18 gRNAs. Here, we demonstrate that PARA is capable of the efficient assembly of transfer RNA/Csy4/ribozyme-based gRNA arrays. To aid in this process and to streamline vector construction, we developed a user-friendly PARAweb tool for designing PCR primers and component DNA parts and simulating assembled gRNA arrays and vector sequences.
- Subjects
GOLDEN Gate Bridge (San Francisco, Calif.); CRISPRS; DNA primers; GENOME editing
- Publication
Cells (2073-4409), 2022, Vol 11, Issue 16, p2467
- ISSN
2073-4409
- Publication type
Article
- DOI
10.3390/cells11162467