CAS9 EFFECTOR-MEDIATED REGULATION OF TRANSCRIPTION, DIFFERENTIATION AND GENE EDITING/LABELING

摘要

The present disclosure relates to methods of and systems for modifying the transcriptional regulation of stem or progenitor cells to promote their differentiation or reprogramming of somatic cells. Further, the labeling and editing of human genomic loci in live cells with three orthogonal CRISPR/Cas9 components allow multicolor detection of genomic loci with high spatial resolution, which provides an avenue for barcoding elements of the human genome in the living state.

主权项

1. A method, comprising:
a) providing;
i) at least one stem cell comprising at least one specific genomic target; andii) a first and second lentiviral vectors, wherein said first lentiviral vector encodes a nuclease deficient Cas9-effector domain fusion protein and said second lentiviral vector comprises at least one sgRNA gene complementary with said specific genomic sequence; b) expressing said first and second lentiviral vectors in said at least one stem cell wherein a nuclease deficient Cas9-effector domain fusion protein/sgRNA complex is formed; and c) regulating transcription of said specific genomic target with said nuclease deficient Cas9-effector domain/sgRNA complex.