Skip to main content

Endogenous Deep Mutational Scans (EDMS)

Command Line Interface

edms -h # or edms <TAB>

Package Organization

  • gen: input/output, data wrangling, generating plots, and statistics.
    import edms.gen.cli as cli
    import edms.gen.image as im
    import edms.gen.io as io
    import edms.gen.plot as p
    import edms.gen.stat as st
    import edms.gen.tidy as t
    
  • bio: molecular biology & tissue culture workflows.
    import edms.bio.clone as cl
    import edms.bio.fastq as fq
    import edms.bio.genbank as gb
    import edms.bio.ngs as ngs
    import edms.bio.pe as pe
    import edms.bio.pegLIT as pegLIT
    import edms.bio.primedesign as primedesign
    import edms.bio.qPCR as qPCR
    import edms.bio.sanger as sanger
    import edms.bio.signature as signature
    import edms.bio.transfect as tf
    
  • dat: interacting with databases.
    import edms.dat.cosmic as co
    import edms.dat.cvar as cv
    import edms.dat.ncbi as ncbi
    

Instructions

Install

  1. Download Anaconda:
  2. Download Git: https://github.com/git-guides/install-git
  3. Clone edms from github:
    cd ~
    mkdir git
    cd git
    git clone https://github.com/marczepeda/edms.git
    cd edms 
    
  4. Make the environment and install edms:
    conda env create -f edms.yml # When conda asks you to proceed, type "y"
    conda activate edms
    pip install -e . # Include the "."
    bash autocomplete.sh # Optional: follow CLI instructions
    conda deactivate
    
  5. Optional: fastq.py UMI methods need umi_tools, cutadapt, samtools, bowtie2, and fgbio in a seperate environment
    conda create -n umi_tools umi_tools cutadapt samtools bowtie2 fgbio
    

Update

  1. Enter the environment and uninstall edms:
    cd ~/git/edms
    conda activate edms
    pip uninstall -y edms
    rm -rf build/ dist/ *.egg-info
    
  2. Pull latest version from github and install edms:
    git pull origin main
    pip install -e . # Include the "."
    conda deactivate
    

PE Strategies

Strategy Description Reference
PE1 Cas9(H840A) - M-MLV RT
+ pegRNA
Search-and-replace genome editing without double-strand breaks or donor DNA
PE2 Cas9(H840A) – M-MLV RT(D200N/L603W/T330P/T306K/W313F)
+ pegRNA
Search-and-replace genome editing without double-strand breaks or donor DNA
PE3 Cas9(H840A) – M-MLV RT(D200N/L603W/T330P/T306K/W313F)
+ ngRNA (targets non-edited strand)
Search-and-replace genome editing without double-strand breaks or donor DNA
PE4 Cas9(H840A) – M-MLV RT(D200N/L603W/T330P/T306K/W313F)
+ MLH1dn (MMR evasion)
Enhanced prime editing systems by manipulating cellular determinants of editing outcomes
PE5 Cas9(H840A) – M-MLV RT(D200N/L603W/T330P/T306K/W313F)
+ MLH1dn (MMR evasion)
+ ngRNA (targets non-edited strand)
Enhanced prime editing systems by manipulating cellular determinants of editing outcomes
PE6a-d Cas9(H840A) – ...
PEa: ... - evo-Ec48 RT
PEb: ... - evo-Tf1 RT
PEc: ... - Tf1 RT variant
PEd: ... - M-MLV RT variant
Phage-assisted evolution and protein engineering yield compact, efficient prime editors
PE6e-f Cas9(H840A) variants – ...
M-MLV RT(ΔRNAseH)
Phage-assisted evolution and protein engineering yield compact, efficient prime editors
PE7 Cas9(H840A) – M-MLV RT(D200N/L603W/T330P/T306K/W313F) - La (RNA binding protein that stabilizes pegRNA)
+/- ngRNA (targets non-edited strand)
Improving prime editing with an endogenous small RNA-binding protein
PEmax Mammalian codon-optimized PE Enhanced prime editing systems by manipulating cellular determinants of editing outcomes
pegRNA spacer - scaffold - RTT - PBS (makes the edit) Search-and-replace genome editing without double-strand breaks or donor DNA
epegRNA spacer - scaffold - RTT - PBS - linker - tevoPreQ (makes the edit; more stable pegRNA) Engineered pegRNAs improve prime editing efficiency
ngRNA spacer - scaffold (targets non-edited strand) Search-and-replace genome editing without double-strand breaks or donor DNA
MLH1dn Dominant negative MLH1 (MMR evasion) Enhanced prime editing systems by manipulating cellular determinants of editing outcomes
silent mutations Larger prime edits are more efficient through bypassing MMR Enhanced prime editing systems by manipulating cellular determinants of editing outcomes
La Small RNA binding protein that stabilizes pegRNA Improving prime editing with an endogenous small RNA-binding protein
PE-eVLP Engineered Virus-Like Particle for Prime Editors Engineered virus-like particles for transient delivery of prime editor ribonucleoprotein complexes in vivo
dNTPs HSCs have low dNTP levels, limiting reverse transcription Enhancing prime editing in hematopoietic stem and progenitor cells by modulating nucleotide metabolism
Vpx HSCs express SAMHD1 (triphosphohydrolase), which depletes dNTPs. Accessory lentiviral protein Vpx, encoded by HIV-2 and simian immunodeficiency viruses (SIVs), associates with the CRL4-DCAF1 E3 ubiquitin ligase to target SAMHD1 for proteasomal degradation. Enhancing prime editing in hematopoietic stem and progenitor cells by modulating nucleotide metabolism
MLH-SB Small protein binder that disrupts MLH1 & PMS2 binding (MMR evasion) AI-generated small binder improves prime editing (Preprint)

Metadata

Release files for edms 0.0.post176

For a detailed explanation of source distributions (sdists) and built distributions (wheels), please see the package formats documentation.

Source distribution (sdist)

Source distribution for edms 0.0.post176
File Size Uploaded
edms-0.0.post176.tar.gz 361.3 kB Details

Built distribution (wheel)

Table of built distributions (wheels) for edms 0.0.post176
File Interpreter ABI Platform
edms-0.0.post176-py3-none-any.whl Python 3 none any Details

Total release size: 731.6 kB

Release files / edms-0.0.post176.tar.gz

Download URL edms-0.0.post176.tar.gz
Size 361.3 kB
Tags Source
SHA-256 checksum
How to use checksums
c4dfb59238bba64825f2c709983bf60a8954d1c726b2bd150a06c46636beaf71
BLAKE2b-256 checksum
How to use checksums
bd147fe661861527557dbb8001bba39e53d7e0f331a19f9f3e812d5e9c0fa3bd
Upload date
Uploaded using Trusted Publishing?
What is trusted publishing?
No
Uploaded via twine/6.1.0 CPython/3.12.2

Release files / edms-0.0.post176-py3-none-any.whl

Download URL edms-0.0.post176-py3-none-any.whl
Size 370.3 kB
Tags Python 3
SHA-256 checksum
How to use checksums
6086fbd52ccc1c515bcc0e49cebd09469044ec5feed953946f9ba175c9407211
BLAKE2b-256 checksum
How to use checksums
432af59b2e751723517e3326d49175043dccbf65734951012b8206beab744979
Upload date
Uploaded using Trusted Publishing?
What is trusted publishing?
No
Uploaded via twine/6.1.0 CPython/3.12.2
Anthropic, PBC Visionary sponsor Bloomberg Visionary sponsor Hudson River Trading Visionary sponsor Meta Visionary sponsor NVIDIA Visionary sponsor Microsoft Sustainability sponsor Depot Continuous Integration AWS Cloud computing and Security Sponsor Datadog Monitoring Fastly CDN Google Download Analytics Sentry Error logging StatusPage Status page