RNA

While incredibly powerful in yeast, genetic analysis of essential human protein complexes/pathways has not been possible until recently.  Now, with the advent of CRISPR-based tools such as base editors, investigating essential processes in human cells is now possible because informative viable missense alleles can be selected, offering exciting possibilities for obtaining new insights.  In ongoing studies, we are deploying cytosine-specific CRISPR base editors in haploid human cells to target the most complex gene expression machine, the human spliceosome, which is considerably more complex in humans than in yeast.  We wish to resolve the paradox of how the human spliceosome maintains fidelity despite the need to be sufficiently flexible to allow regulated splicing.  We are also curious about how transcription and RNA processing events may be coupled in vivo.