sgrna_designer

Python library to design sgRNAs for CRISPR tiling screens

The primary function of this package is design_sgrna_tiling_library, in which you can input a list of ensembl transcript IDs, specify a region of interest (e.g. three_prime_UTR) and get all sgRNAs tiling those transcript regions.

Install

pip install git+https://github.com/gpp-rnd/sgrna_designer.git#egg=sgrna_designer

An example

In this example we'll design sgRNAs tiling the 3' UTR of PDL1 (CD274) and BRAF

Note: You must also have pandas installed to run this tutorial

from sgrna_designer.design import design_sgrna_tiling_library

target_transcripts = ['ENST00000381577', 'ENST00000644969'] # [PDL1, BRAF]

Note the design function is agnostic to CRISPR enzyme and pam preferences, so you must specifiy the following parameters in a design run:

• region: broad region you are trying to target (e.g. UTR)
• region: more specific region you are trying to target (e.g. three_prime_UTR)
• expand_3prime: amount to expand region in 3' direction
• expand_5prime: amount to expand region in 5' direction
• context_len: length of context sequence
• pam_start: position of PAM start relative to the context sequence
• pam_len: length of PAM
• sgrna_start: position of sgRNA relative to context sequence
• sgrna_len: length of sgRNA sequence
• pams: PAMs to target
• sg_positions: positions within the sgRNA to annotate and target (e.g. [4,8] for nucleotides 4 and 8 of the sgRNA for a base editing window)

sgrna_designs = design_sgrna_tiling_library(target_transcripts, region_parent='UTR',
                                            region='three_prime_UTR', expand_3prime=30,
                                            expand_5prime=30, context_len=30, pam_start=-6,
                                            pam_len=3, sgrna_start=4, sgrna_len=20,
                                            pams=['AGG', 'CGG', 'TGG', 'GGG'],
                                            sg_positions=[4, 8], flag_seqs=['TTTT', 'CGTCTC', 'GAGACG'],
                                            flag_seqs_start=['TCTC', 'AGACG'], flag_seqs_end=['GAGAC'])
sgrna_designs

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