‘Mini’ U6 Pol III promoter exhibits nucleosome redundancy and supports multiplexed coupling of CRISPR/Cas9 effects

RNA polymerase III (Pol III) promoters express short non-coding RNAs and have been adopted for expression of microRNA, interference RNA, and CRISPR single guide RNA (sgRNA). Vectors incorporating H1 and U6 Pol III promoters are being applied for therapeutic genome editing, including multiplexed CRIS...

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Bibliographic Details
Published in:Gene therapy 2020-09, Vol.27 (9), p.451-458
Main Authors: Preece, Roland, Georgiadis, Christos, Gkazi, Soragia Athina, Etuk, Aniekan, Christi, Abraham, Qasim, Waseem
Format: Article
Language:English
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Biomedical and Life Sciences
Biomedical engineering
Biomedicine
Brief Communication
Cell Biology
CRISPR
CRISPR-Cas Systems
DNA-directed RNA polymerase
Expression vectors
Gene Editing
Gene Expression
Gene Therapy
Genetic transcription
Genome editing
Genomes
Genomics
Health aspects
Histocompatibility antigen HLA
Histocompatibility antigens
HLA histocompatibility antigens
Human Genetics
Humans
Long terminal repeat
Lymphocytes T
Medical research
Medicine, Experimental
MicroRNA
miRNA
Nanotechnology
Non-coding RNA
Nucleosomes - genetics
Promoter Regions, Genetic
Promoters
RNA polymerase
RNA polymerases
RNA, Guide, CRISPR-Cas Systems
RNA-mediated interference
T cell receptors
T cells
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Summary:RNA polymerase III (Pol III) promoters express short non-coding RNAs and have been adopted for expression of microRNA, interference RNA, and CRISPR single guide RNA (sgRNA). Vectors incorporating H1 and U6 Pol III promoters are being applied for therapeutic genome editing, including multiplexed CRISPR/Cas9 effects. We report a nucleosome-depleted, minimal U6 promoter, which when embedded within lentiviral long terminal repeat (LTR) regions, supports high level transcriptional activity. Furthermore, duplex minimal H1 & U6 promoters transcribed dual sgRNAs for simultaneous disruption of T cell receptor (TCR) and human leukocyte antigen (HLA) molecules, supporting efficient generation of ‘universal’ CAR T cells.
ISSN:0969-7128
1476-5462
DOI:10.1038/s41434-020-0142-z