Lessons Learned from Neutralizing Drives

Table of Contents Below. Dr. Ethan Bier describes research and development efforts focused on technologies that might be useful in halting and reversing the spread of engineered gene drives. 00:00 Speaker Introduction (David O’Brochta) 1:22 Introductory Remarks 1:55 Efficient drive at the kh locus 3:13 Maternal inheritance of Cas9 3:58 kh-2 recoded drive 5:50 kh-1 drive in cage experiments – drives poorly 6:38 kh-2 drive in cage experiments – drives well 8:40 Intro to mitigation strategies 10:13 Allelic drive strategy- 11:30 Example Reversing DDT resistance 13:20 Allelic drive description 13:57 Results 15:58 ERACRs: Delete and replace gene drive - description 17:42 Model kinetics 18:15 Examples of various ERACRs – 20:10 Genetic analysis of process – detailed genetics with ERACR-2 Initial cross 23:26 Comparing ERACRs 24:49 Lesson 1 Marking donor vs receiver chromosome is essential 25:34 Lesson 2 ERACRs are not perfect 26:10 Lesson 3 ERACRs can damage target chromosome -Deletion without copying 27:57 Lesson 4 Single sided homology leads to chromosome damage Single cut vs double cut ERACRs 31:56 Lesson 5 ERACRs generate rare rearrangements. 36:07 Lesson 6 ERACRs can drive very efficiently Drive vs ERACRs 39:20 eCHACR description 42:43 Lesson 7 eCHACR copy very efficiently 45:54 Lesson 8 eCHACR reduce drive 47:27 Lesson 9 eCHACR can damage gene drive chromosome 48:26 eCHACR cutting a chromosome twice bias inheritance of donor chromosomes 50:35 Lesson 10 eCHACR drive efficiently in cages. 53:05 Summary of Lessons 56:33 Remaining Questions 59:52 Future Goals 1:01:21 Acknowledgements 1:03:37 Questions and Answers