Reorganize repository structure and update documentation for GitHub integration

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james-m-jordan 2025-05-06 09:33:40 -04:00
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This file will be updated with a summary of major actions, sessions, and changes as the system is used.
## [2025-05-06] Ybx1 mRNA Stability Project
- Created new project entry for Post-transcriptional regulation by Ybx1
- Created subproject for mRNA stability measurements
- Created new protocol for Ybx1 knockdown mRNA stability assay (PROT-0035)
- Generated experiment YAML for initial Ybx1 knockdown study (EXP-0225)
- Created data directories for experiment results
- Created GitHub issues with unique hash identifiers for experiment tracking:
- RNA collection for Ybx1 knockdown experiment (fd80f1a)
- Verify Ybx1 knockdown by qPCR (fd80f2b)
- Actinomycin D time course (fd80f3c)
- RNA extraction, cDNA synthesis, qPCR (fd80f4d)
## [Unreleased]
- Initial scaffolding and setup.

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title: mRNA stability measurement after Ybx1 knockdown
description: Investigation of mRNA stability changes in selected genes after Ybx1 knockdown, using siRNA and actinomycin D to assess post-transcriptional regulation
date_started: 2025-05-06
lead: Dr. Jim Jordan
team_members:
- Dr. Jim Jordan
- Jack Zhao
parent_project: Post-transcriptional regulation by Ybx1
associated_protocols:
- Ybx1 knockdown mRNA stability assay
experiments:
- TBD
notes: |
This subproject aims to determine if Ybx1 knockdown influences the stability of mRNAs,
particularly those with known or suspected post-transcriptional modifications (m5C or m6A).
Specific aims:
1. Establish efficient Ybx1 knockdown using siRNA
2. Measure half-lives of selected target mRNAs in control vs. Ybx1 knockdown conditions
3. Identify genes most affected by Ybx1 depletion
4. Correlate mRNA stability changes with RNA modifications if possible
Initial target genes include Myc, p53, and other transcripts with known post-transcriptional regulation.
This experiment will employ actinomycin D to block transcription, followed by time-course
sampling and RT-qPCR to measure the decay rates of specific mRNAs.

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_Note: Burst 5 implemented Smart-Fill using ChromaDB and OpenAI embeddings to suggest metadata fields for protocols and experiments based on repository content. Next, GitHub integration (commit, push, issues, PRs) will be developed._
## Burst 6: GitHub Integration
- [ ] Implement commit and push logic
- [ ] Implement GitHub issue creation
- [x] Implement commit and push logic
- [x] Implement GitHub issue creation with short hash identifiers
- [ ] Implement draft pull request creation
- [ ] Add structured commit messages and PR descriptions
- [ ] Begin repository structure reorganization
_Note: Burst 6 implemented basic GitHub integration including commit/push functionality and issue creation with unique hash identifiers for better traceability. Work has begun on converting projects to aims and subprojects to projects for better GitHub issue organization._
## Burst 7: Ybx1 Knockdown mRNA Stability Experiment
- [x] Create project for Post-transcriptional regulation by Ybx1
- [x] Create subproject for mRNA stability measurements
- [x] Create protocol for Ybx1 knockdown mRNA stability assay (PROT-0035)
- [x] Generate experiment YAML for initial Ybx1 knockdown study (EXP-0225)
- [x] Create data directories for experiment results
- [x] Create morning push checklist (2025-05-06)
- [x] Create GitHub issues with unique hash identifiers for experiment tracking
_Note: Burst 7 implemented the first real-world experiment using the system, creating all necessary files, issues, and documentation with the new hash identifier system for better future-proofing._
## Burst 8: Repository Restructuring for GitHub Integration
- [ ] Convert "projects" to "aims" for higher-level organization
- [ ] Convert "subprojects" to "projects" for better GitHub issue integration
- [ ] Update all existing YAML files to reflect new structure
- [ ] Update documentation to reflect new naming conventions
- [ ] Create migration guide for users
_Note: This burst focuses on reorganizing the repository structure to better integrate with GitHub project management features, enabling more intuitive issue organization and tracking._
## Burst 9: Experiment Update, Validation, and Logging
- [x] Implement experiment update logic (handle_update_experiment)

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_Note: This file is now automatically updated by the agent/task-runner. Tasks are marked done when experiments are completed, and new tasks may be added as needed._
## Lab Tasks
- [ ] (Add lab tasks here)
### Ybx1 Knockdown mRNA Stability Experiment (EXP-0225)
- [ ] Day 1 (May 6, 2025): Complete reverse transfection setup
- [ ] Thaw HEK293T cells and check viability
- [ ] Prepare transfection master mixes (siRNA targeting Ybx1 and Control siRNA)
- [ ] Set up reverse transfection in 24-well plates
- [ ] Update experiment status in repository
- [ ] Day 3 (May 8, 2025): Verify Ybx1 knockdown by qPCR
- [ ] Day 3 (May 8, 2025): Begin actinomycin D time course experiment
- [ ] Collect t=0 samples before actinomycin D addition
- [ ] Add actinomycin D (5 μg/ml) to remaining wells
- [ ] Collect samples at 2h, 4h, and 8h timepoints
- [ ] Day 4 (May 9, 2025): Complete RNA extractions
- [ ] Day 4 (May 9, 2025): Synthesize cDNA and perform qPCR
- [ ] Submit final data to repository and link to experimental record
### Materials and Resources
- [ ] Confirm Actinomycin D stock availability
- [ ] Ensure sufficient RNA extraction reagents are available
- [ ] Check qPCR primer stocks for all target genes
## Development Tasks
- [ ] (Add development tasks here)
- [ ] Reorganize repository structure: convert "projects" to "aims" and "subprojects" to "projects" for better GitHub integration
- [ ] Improve GitHub issue integration with short hash identifiers for future-proofing
- [ ] Update documentation to reflect new naming conventions
- [ ] Evaluate need for TASKS.md file in the new structure (consider GitHub issues as primary task tracker)
_This file will be updated as tasks are added or completed during implementation._

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