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New Strategies for Forward Genetic Screening and Single Cell Analysis with CRISPRi Transcriptional Repressors
PRESENTER:
Dr. Tiffany Bernardo | Research Technology Specialist, Advanced Genomics and Cell Culture Models at MilliporeSigma
PRESENTER'S BIO:
Tiffany is the Research Technology Specialist for the Southeast and Mid-Atlantic US covering advanced cell culture applications, functional genomics, and protein platforms. She completed her PhD. in Developmental Biology and Molecular Genetics from the University of Pittsburgh and brings extensive knowledge of molecular, developmental, and stem cell biology, as well as substantial teaching experience.
ABSTRACT:
Pairing CRISPR-based genetic screens with single-cell transcriptional phenotypes have advanced efforts to explore the function of mammalian genes and genetic networks. Our CRISPRi technology utilizes catalytically inactive KRAB-dCas9 along with a single guide RNA (sgRNA) to allow inhibition at the endogenous DNA level. Here, we demonstrate that improved sgRNA scaffold increases knockdown efficiency in difficult to repress targets as well as CRISPRi’s utility with the 10x Genomics Feature Barcoding platform for single-cell analysis.
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