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PARA: A New Platform for the Rapid Assembly of gRNA Arrays for Multiplexed CRISPR Technologies.

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.

SUBMITTER: Yuan G

PROVIDER: S-EPMC9406951 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature

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PARA: A New Platform for the Rapid Assembly of gRNA Arrays for Multiplexed CRISPR Technologies.

Yuan Guoliang G Martin Stanton S Hassan Md Mahmudul MM Tuskan Gerald A GA Yang Xiaohan X

Cells 20220809 16

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 proces ...[more]

PMID: 36010544

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