BIOL 330 Lecture Notes - Lecture 12: Riboswitch, Cas9, Crispr
Document Summary
Rna functions as a regulator by forming a region of secondary structure (either inter- or intramolecular) that changes the properties of a target sequence. Knocking out mammalian genes with crispr: express cas9 and single guide rna *which matches a gene of interest) in mammalian cells, to generate a targeted double stranded break. Non- homologous end joining with fix it, usually inserting or deleting a few bases, often inactivating the gene. Over expressing human genes with crispr: fused a transcription factor to a defective cas9 that cannot cleave dna, transduced this along with a grna library, the grna binds upstream of every gene in the genome. 23 nucleotide rnai molecules: risc- rna induced silencing complex, a ribonucleoprotein particle composed of a short, single-stranded sirna and a nuclease that cleaves mrnas complementary to the sirna. It receives sirna from dicer and delivers it to the mrna.
