Skip to content
New issue

Have a question about this project? Sign up for a free GitHub account to open an issue and contact its maintainers and the community.

By clicking “Sign up for GitHub”, you agree to our terms of service and privacy statement. We’ll occasionally send you account related emails.

Already on GitHub? Sign in to your account

Inferelator workflow for subsetted Seurat .h5ad file #64

Open
jsacco1 opened this issue Jun 22, 2023 · 1 comment
Open

Inferelator workflow for subsetted Seurat .h5ad file #64

jsacco1 opened this issue Jun 22, 2023 · 1 comment

Comments

@jsacco1
Copy link

jsacco1 commented Jun 22, 2023

I have a workflow question. Here is some background:

I have a Seurat RDS file of integrated ATAC-seq and RNA-seq data from human samples. This data measures TF expression. After running PCA and UMAP methods, I end up with numbered clusters, one of which corresponds to a strongly expressed TF (that is,
strong enough that a cluster number can be labeled with that TF). I want to run Inferelator 3.0 on cells from that TF cluster. I subsetted the Seurat object first by cluster number, and then by expression level (> 0.5).

Question:
I converted this new, smaller Seurat object of ~200 cells into loom file, then into a h5ad file. How do I run Inferelator 3.0 on this .h5ad file?

Also: I have publicly available bulk ATAC-seq data, with which I ran Inferelator Prior to make the priors file. Would it be better to use a experimental ATAC-seq, although it would have lower depth?

@asistradition
Copy link
Collaborator

You can read in h5ad files with the following (change paths to match your configuration):

worker = inferelator_workflow(regression='stars', workflow='single-cell')
worker.set_file_paths(
        input_dir='.',
        output_dir='.',
        priors_file='priors.tsv',
        gold_standard_file='gold_standard.tsv'
)
worker.set_expression_file(
    h5ad='data.h5ad',
    h5_layer=None
)

h5_layer=None uses the adata.X array, if you pass an argument it'll use adata.layers[h5_layer] instead.

As for the best ATAC data to use for any project, it's so specific to the data that I don't think I can give you any useful general advice.

Sign up for free to join this conversation on GitHub. Already have an account? Sign in to comment
Labels
None yet
Projects
None yet
Development

No branches or pull requests

2 participants