CMMB 403 final Exam New Year 2024| Questions and Answers| Certified Quality Graded A+

CMMB 403 final Exam New Year 2024| Questions and Answers| Certified Quality Graded A+ what does CRISPR use in order to generate changes in genome? is CRISPR forward or reverse genetics? - nuclease - 20 nucleotide RNA sequence to target specific sites in the genome - reverse genetics what is the role of Cas9 in CRISPR endonuclease that catalyzes double stranded breaks what are the two repair paths cells can use to repair double stranded breaks from CRISPR-Cas9 1. non-homologous end joining (NHEJ) - naturally try to repair - imperfect, creates indels causing frameshift mutation 2. directed homology repair - sequence homology on 5' and 3' ends to encourage homologous recombination flanking the double stranded breaks - insert known sequences to try and treat genetic diseases explain the method of conditional gene inactivation - make targetting construct in lab - electroporate construct into embryonic stem cells to target endogenous Shox2 locus - mice generated by blastocyst injection into ESCs carry loxP sites therefore can delete Shox2