Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Cell line (Homo sapiens) | HEK293T | ATCC | ATCC:CRL-3216 | |
| Recombinant DNA reagent | pCMV-Rep78/68 plasmid library | This paper | RRID:Addgene_198050 | Complete list of plasmid sequences included in library can be found in the GitHub repo linked below |
| Recombinant DNA reagent | pEf1a-Luc-WPRE (ssAAV) plasmid | Addgene | RRID:Addgene_87951 | |
| Recombinant DNA reagent | pCAG-Luc (ssAAV) plasmid | Addgene | RRID:Addgene_83281 | |
| Recombinant DNA reagent | pCAG-GFP (scAAV) plasmid | Addgene | RRID:Addgene_83279 | |
| Sequence-based reagent | 5′-GAGCATCTGCCCGGCATTTC-3′ | This paper | Forward primer, binds to 5′ region of AAV2 rep gene | |
| Sequence-based reagent | 5′-ATCTGGCGGCAACTCCCATT-3′ | This paper | Reverse primer, binds to 5′ region of AAV2 rep gene | |
| Sequence-based reagent | 5′-HEX-ACAGCTTTG-ZEN- TGAACTGGGTGGCCGA-3IABkFQ-3′ |
This paper | HEX-based probe, binds to 5′ region of AAV2 rep gene | |
| Sequence-based reagent | 5′-GAACCGCATCGAGCTGAA-3′ | This paper | Forward primer, binds to GFP sequence | |
| Sequence-based reagent | 5′-TGCTTGTCGGCCATGATATAG-3′ | This paper | Reverse primer, binds to GFP sequence | |
| Sequence-based reagent | 5′-FAM-ATCGACTTC-ZEN- AAGGAGGACGGCAAC-3IABKFQ-3′ |
This paper | FAM-based probe, binds to GFP sequence | |
| Sequence-based reagent | 5′-CATGTACCGCTTCGAGGAG-3′ | This paper | Forward primer, binds to luciferase sequence | |
| Sequence-based reagent | 5′-GAAGCTAAATAGTGTGGGCACC3′ | This paper | Reverse primer, binds to luciferase sequence | |
| Sequence-based reagent | 5′-FAM-CTTGCGCAG-ZEN- CTTGCAAGACTATAAGATTC-3IABKFQ-3′ |
This paper | FAM-based probe, binds to luciferase sequence | |
| Antibody | Anti-Rep 303.9 (mouse monoclonal) | American Research Products | American Research Products: 03-61069; RRID:AB_1540388 | 1:100 |
| Antibody | B1 anti-VP (mouse monoclonal) | American Research Products | American Research Products: 03-61058; RRID:AB_1540385 | 1:250 |
| Antibody | Anti-β-actin (rabbit polyclonal) | Abcam | Abcam: ab8227; RRID:AB_2305186 | 1:20,000 |
| Antibody | Anti-mouse IgG IRDye 800CW (goat monoclonal) | LI-COR | LI-COR: 925-32210; RRID:AB_2687825 | 1:10,000 |
| Antibody | Anti-rabbit IgG IRDye 680RD (donkey monoclonal) | LI-COR | LI-COR: 925-68073; RRID:AB_2716687 | 1:10,000 |
| Commercial assay or kit | Promega One-Glo EX Luciferase Assay System | Promega | Promega:E8110 | |
| Commercial assay or kit | Capsid ELISA | Progen | Progen:PRATV | |
| Software, algorithm | Python scripts for library design and data analysis | This paper | https://github.com/churchlab/aav_rep_scan; churchlab, 2023 |