منابع مشابه
Genome-scale CRISPR pooled screens.
Genome editing technologies such as clustered regularly interspaced short palindromic repeats (CRISPR) systems have ushered in a new era of targeted DNA manipulation. The easy programmability of CRISPR using short oligonucleotides enables rapid synthesis of large-scale libraries for functional genetic screens. Here we present fundamental concepts and methods for pooled CRISPR screens and review...
متن کاملEvaluation and Design of Genome-Wide CRISPR/SpCas9 Knockout Screens
The adaptation of CRISPR/SpCas9 technology to mammalian cell lines is transforming the study of human functional genomics. Pooled libraries of CRISPR guide RNAs (gRNAs) targeting human protein-coding genes and encoded in viral vectors have been used to systematically create gene knockouts in a variety of human cancer and immortalized cell lines, in an effort to identify whether these knockouts ...
متن کاملDissecting Immune Circuits by Linking CRISPR-Pooled Screens with Single-Cell RNA-Seq
In multicellular organisms, dedicated regulatory circuits control cell type diversity and responses. The crosstalk and redundancies within these circuits and substantial cellular heterogeneity pose a major research challenge. Here, we present CRISP-seq, an integrated method for massively parallel single-cell RNA sequencing (RNA-seq) and clustered regularly interspaced short palindromic repeats ...
متن کاملPICKLES: the database of pooled in-vitro CRISPR knockout library essentiality screens
The adaptation of CRISPR/Cas9 systems for pooled library genetic knockout screens in mammalian cells has substantially advanced the state of the art in human functional genomics. Screening panels of cell lines for genes whose knockout imposes a significant fitness defect has dramatically expanded our catalog of high-confidence essential genes, and has already proven useful in identifying tumor-...
متن کاملDetermination of sample size in genome-scale RNAi screens
MOTIVATION For genome-scale RNAi research, it is critical to investigate sample size required for the achievement of reasonably low false negative rate (FNR) and false positive rate. RESULTS The analysis in this article reveals that current design of sample size contributes to the occurrence of low signal-to-noise ratio in genome-scale RNAi projects. The analysis suggests that (i) an arrangem...
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ژورنال
عنوان ژورنال: Analytical Biochemistry
سال: 2017
ISSN: 0003-2697
DOI: 10.1016/j.ab.2016.05.014