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Exam (elaborations)

Module 6: Genetic Engineering Exam Questions With Verified Answers.

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Module 6: Genetic Engineering Exam Questions With Verified Answers. How are ds breaks repaired - answernon-homologous end joining (NHEJ) homologous directed recombination (HDR) non-homologous end joining (NHEJ) definition - answerligated 2 broken ends together (not template repaired) typically results in insertions or deletion homologous directed recombination (HDR) definition - answertemplate dependent pathway for DSB repair can insert donor molecule at target types of application of gene selection - answerNatural selection Human intervention example of natural gene selection - answerDarwin's finches example of gene selection via human intervention - answermodern corn grain modified by humans what is genetic engineering - answerthe deliberate modification of the characteristics of an organism by manipulating its genetic material integrating genetic info from one species to another ex vivo gene editing process - answerremove cells correct gene autologous transplantation (back into same patient) injection of vectors with therapeutic elements difficulties of ex vivo studies - answerdifficult to ensure cell survival after removal (need to know exactly what to supplement) gene editing process for cancer immunotherapy - answercollect WBC isolate T cells engineer T cells to express CARs that recognize cancer cells culture modified T cells infuse cell into patient (modified T cells track down and kill cancer) types of genome editing - answergermline and somatic genome editing germline mutation - answermutation that occur in gametes can be passed to offspring somatic mutation - answera mutation that occurs in a single body cell tissues derived from the cell are affected mutation not inherited forward genetics - answerphenotype to gene begin with phenotype and then determine a gene that encodes the phenotype compare genetic of those with and without the given phenotype reverse genetics - answergene to phenotype have gene, but don't know its function induce gene and observe to identify phenotype example of reverse genetics - answerinject neu transgene in mice pros of drugs and radiation to induce mutations - answeruseful for large complex genetic mutations (translocations) cons of drugs and radiation to induce mutations - answerrandom mutations occur cannot control who and when were zinc finger nucleases discovered - answerHolmes, 2005 who and when were TALENs discovered - answervoytas, 2010 who and when were CRISPR discovered - answerCharpentier and Doudna 2012 methods to edit genes - answerZFNs TALENs CRISPR/Cas9 what can ZFNs, TALENs and CRISPR/Cas9 all do - answercan induce double stranded breaks at targeted sites Zinc finger nucleases (ZFNs) function - answerzinc fingers fit into the major groove of DNA, recognizing specific sequences induces break at fok1 cleavage domain transcription activator-like effector nuclease (TALENs) function - answerengineered restriction enzymes central repeat domain, have repeating units (33-35 aa), repeats similar except for 2 variable aa (repeat variable di-resides) TALEN recognizes single nucleotide based on RVDs how long does it take to develop 1 TALEN pair - answer fok1 - answerA non-specific restriction enzyme that can only function as a dimer. Can cut anywhere in DNA.

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