Journal: eLife

Article Title: Systematic lncRNA mapping to genome-wide co-essential modules uncovers cancer dependency on uncharacterized lncRNAs

doi: 10.7554/eLife.77357

Figure Lengend Snippet: ( A ) Density plots showing the distribution of proliferation-inducing and -suppressing lncRNA levels in cells treated with doxorubicin (Dox), 5-fluorouracil (5-FU), or ionizing radiation (IR) in seven bulk RNA-seq data and seven bulk RNA-seq data of cells treated with Nutlin-3. ( B ) Density plots as in ( A ) from 24 single-cell RNA-seq data from cells treated with Nutlin-3 (7 p53 WT and 17 p53 MUT cell lines). ( C ) Heatmap showing fold change of predicted proliferation-inducing and -suppressing lncRNAs across 14 RNA-seq datasets, indicated in ( A ). n is the number of consistently upregulated lncRNAs across the RNA-seq datasets with statistical significance (Benjamini–Hochberg [BH] adjusted *p<0.05) calculated by the meta-analysis tool RobustRankAggreg; a subset of the top proliferation suppressors are highlighted in the box. ( D ) The number of ChIP-seq datasets showing p53-binding sites near the transcription start site of the 13 meta-analyses-derived significantly upregulated lncRNAs, indicated in ( C ), or background lncRNAs. For ( A , B , D ), p-values were calculated from two-tailed Wilcoxon rank-sum tests. ( E ) The expression fold change of two poorly characterized top predicted proliferation-suppressing lncRNAs in cells treated with p53-activating agents (see A ) compared to control cells. The X-axis indicates the accession number of the 14 RNA-seq datasets present in the Gene Expression Omnibus database. ( F ) Two lung adenocarcinoma (LUAD) cell lines treated with 10 µM of Nutlin (NUT), etoposide (ETO), or cisplatin (CIS) or received 5 Gy of gamma-radiation (IR) were harvested at intervals. qRT-PCR (triplicate) for specific lncRNA was performed. BAX and AchR mRNA are positive and negative controls, respectively. RNA levels were normalized to β-ACTIN, and values are presented as 2 -ΔΔCt , with vehicle control-treated cells set at 1 (black line); data are mean ± SEM. For A549, PSLR-1 *p<6.86 × 10 –4 , PSLR-2 *p<1.17 × 10 –2 , BAX *p<4.93 × 10 –6 ; for H460, PSLR-1 *p<6.95 × 10 –6 , PSLR-2 *p<5.69 × 10 –3 , and BAX *p<3.99 × 10 –4 ; two-tailed t -tests. Figure 3—source data 1. qRT-PCR for .

Article Snippet: Transfected construct ( Homo sapiens ) , PSLR-1 , VectorBuilder , FAM198B-AS1 VB200405-4575kbk , Lentiviral construct to express the lncRNA.

Techniques: RNA Sequencing, ChIP-sequencing, Binding Assay, Derivative Assay, Two Tailed Test, Expressing, Control, Gene Expression, Quantitative RT-PCR

Journal: eLife

Article Title: Systematic lncRNA mapping to genome-wide co-essential modules uncovers cancer dependency on uncharacterized lncRNAs

doi: 10.7554/eLife.77357

Figure Lengend Snippet: ( A ) Expression of PSLR-1 and -2 was measured by qRT-PCR (triplicate) in normal human lung tissue (n = 5) and LUAD patient samples (n = 10). RNA levels were normalized to β-ACTIN and values are presented as 2 -ΔCt ; data are mean ± SEM. PSLR-1, *p=7.75 × 10 –9 ; PSLR-2, *p=2.77 × 10 –8 ; two-tailed t -tests. ( B–D ) RNA-sequencing of A549 LUAD cell line after expression of each of the two long noncoding RNAs (lncRNAs) or vector control (quadruplicate). ( B ) Genes that have significant (adjusted regression p<10 –3 ) positive or negative associations with the indicated lncRNA in TCGA LUAD data were selected. Empirical cumulative distribution of expression fold changes of these genes in A549 cells expressing the indicated lncRNA compared with vector control. ( C ) Empirical cumulative distribution of expression changes of curated tumor-suppressor genes and oncogenes in A549 cells expressing the indicated lncRNA compared with vector control. For ( B , C ), x-axis indicates the expression fold changes of genes, y-axis estimates the percentage of genes at a log2 fold-change value indicated on the x-axis, and p-values were from two-tailed Wilcoxon rank-sum tests. ( D ) Gene Set Enrichment Analysis (GSEA) demonstrated top up- and downregulated Hallmark genesets after expression of the indicated lncRNA in A549 cells compared with vector control. Positive and negative normalized enrichment scores (NES) indicate up- and downregulation, respectively. Significantly enriched (false discovery rate [FDR] < 0.05) proliferation-associated pathways/gene signatures indicated. ( E ) Enrichr-calculated combined score indicates transcription factor (TF) enrichment of genes altered upon expression of the indicated lncRNA in A549 LUAD cells. ( F ) HOMER-calculated promoter motif enrichment of downregulated genes upon indicated lncRNA expression in LUAD cells. CHR motif elements are shown (right). Figure 4—source data 1. qRT-PCR for .

Article Snippet: Transfected construct ( Homo sapiens ) , PSLR-1 , VectorBuilder , FAM198B-AS1 VB200405-4575kbk , Lentiviral construct to express the lncRNA.

Techniques: Expressing, Quantitative RT-PCR, Two Tailed Test, RNA Sequencing, Plasmid Preparation, Control

Journal: eLife

Article Title: Systematic lncRNA mapping to genome-wide co-essential modules uncovers cancer dependency on uncharacterized lncRNAs

doi: 10.7554/eLife.77357

Figure Lengend Snippet: The lncRNAs PSLR-1 , PSLR-2 , or MALAT1 or vector control were expressed (lentivirus) in LUAD cell lines. ( A ) MTT assays (quadruplicate) performed at 24 hr intervals. Each assay performed 2–4 independent times for both cell lines and one representative experiment is shown ± SD; A549 *p=1.11 × 10 –4 and **p<3.03 × 10 –4 , H460 *p=2.04 × 10 –5 and **p<2.28 × 10 –4 ; two-tailed t -tests. ( B ) Colonies from colony formation assays were counted 14 days after lncRNA expression. Assays (triplicate) performed 2–3 independent times and a representative experiment ± SEM and images (no magnification) shown; A549 *p=1.49 × 10 –2 and **p<2.09 × 10 –4 , H460 *p=3.42 × 10 –2 and **p<1.16 × 10 –3 ; two-tailed t -tests. ( C ) Live cell number determined by Trypan Blue dye exclusion 48 hr after placing cells in 6-well plates. Mean of four independent experiments for both cell lines ± SD is graphed; A549 *p=5.77 × 10 –4 and **p<2.68 × 10 –5 , H460 *p=2.90 × 10 –5 and **p<3.63 × 10 –6 ; two-tailed t -tests. ( D ) Cell cycle evaluated at intervals following propidium iodide staining and flow cytometry; representative histograms from one experiment shown (left). Graphs of the cell cycle phases represent the mean of four independent experiments for both cell lines at 48 hr ± SEM; A549 *p=8.82 × 10 –3 and **p<1.73 × 10 –2 , H460 *p=4.00 × 10 –3 and **p<1.35 × 10 –3 ; two-tailed t -tests. Figure 5—source data 1. MTT for . Figure 5—source data 2. Colony number for . Figure 5—source data 3. Live cell number for . Figure 5—source data 4. Cell cycle for .

Article Snippet: Transfected construct ( Homo sapiens ) , PSLR-1 , VectorBuilder , FAM198B-AS1 VB200405-4575kbk , Lentiviral construct to express the lncRNA.

Techniques: Plasmid Preparation, Control, Two Tailed Test, Expressing, Staining, Flow Cytometry

Journal: eLife

Article Title: Systematic lncRNA mapping to genome-wide co-essential modules uncovers cancer dependency on uncharacterized lncRNAs

doi: 10.7554/eLife.77357

Figure Lengend Snippet:

Article Snippet: Transfected construct ( Homo sapiens ) , PSLR-1 , VectorBuilder , FAM198B-AS1 VB200405-4575kbk , Lentiviral construct to express the lncRNA.

Techniques: Transfection, Construct, Plasmid Preparation, Control, Staining, Sequencing, SYBR Green Assay, cDNA Synthesis, Isolation, Proliferation Assay, Colony Assay, Cell Cycle Assay, Positive Control, Software

Journal: iScience

Article Title: Identification of NUV-244 as a PNPLA3 I148M degrading small molecule

doi: 10.1016/j.isci.2025.112384

Figure Lengend Snippet: HTS for the identification of PNPLA3 I148M modulators (A) Huh7 parental control or Huh7 cells expressing PNPLA3 I148M-tagRFP were stained for PNPLA3 by immunofluorescence. In PNPLA3 I148M expressing cells, PNPLA3 I148M-tagRFP is located on the surface of lipid droplets (yellow channel) as seen by antibody staining against PNPLA3 (green channel) and the PNPLA3 I148M-tagRFP fluorescent signal (red channel). Blowup of cell overview pictures (dotted square) show PNPLA3 I148M-tagRFP being located on the surface of lipid droplets. Hoechst staining in blue. Scale bars ∼10μm. Representative images shown from n > 3 independent experiments. (B) Example pictures from Huh7 parental controls (left, not expressing PNPLA3 I148M-tagRFP) showing lipid droplets and no staining in the RFP channel, DMSO treated control of Huh7 cells expressing PNPLA3 I148M-tagRFP with PNPLA3 I148M-tagRFP co-localized to lipid droplets (middle) and a hit from the HTS showing strong reduction of PNPLA3 I148M-tagRFP on lipid droplets. Scale bar ∼10μm. Representative images shown from n > 3 independent experiments.

Article Snippet: Lentiviral constructs for PNPLA3 expression , Sirion Biotech , custom.

Techniques: Control, Expressing, Staining, Immunofluorescence

Journal: iScience

Article Title: Identification of NUV-244 as a PNPLA3 I148M degrading small molecule

doi: 10.1016/j.isci.2025.112384

Figure Lengend Snippet: NUV-244 properties (A) NUV-244 structure and basic EC50 data. The listed properties were calculated with RDKit [RDKit: Open-source cheminformatics. https://www.rdkit.org ] and satisfy Lipinski’s rule of five as well as Veber’s rule. , (B) 1μM NUV-244 reduces PNPLA3 I148M-tagRFP from the surface of lipid droplets after 24h incubation (white arrows). Hoechst in blue, lipid droplets in yellow (upper panel). Hoechst in blue, PNPLA3 I148M-tagRFP in red (lower panel). Scale bar ∼10μm. Representative images shown from n > 3 independent experiments. (C) 1μM NUV-244 shows no effect on PNPLA2 staining on lipid droplets. Hoechst in blue, lipid droplets in yellow (upper panel). Hoechst in blue, PNPLA2 in red (lower panel). Blowup of cell overview pictures (dotted square) show PNPLA2 remaining on the surface of lipid droplets after NUV-244 treatment. Scale bar ∼10μm. Representative images shown from n = 2 independent experiments. (D) Dose-response curves for NUV-244 on Huh7 PNPLA3 I148M-tagRFP expressing cells (black curve) and immunofluorescence against PNPLA2 (red curve). Dose-response with n > 3 per concentration. Quantification of signal intensity on lipid droplets. Curves are normalized to DMSO treated PNPLA3 I148M-tagRFP controls (0) and parental Huh7 cells (not expressing PNPLA3, -100) or in case of PNPLA2 immunofluorescence to DMSO treated (0) and secondary antibody controls (−100). Representative data shown from n > 2 independent experiments.

Article Snippet: Lentiviral constructs for PNPLA3 expression , Sirion Biotech , custom.

Techniques: Incubation, Staining, Expressing, Immunofluorescence, Concentration Assay

Journal: iScience

Article Title: Identification of NUV-244 as a PNPLA3 I148M degrading small molecule

doi: 10.1016/j.isci.2025.112384

Figure Lengend Snippet: Summary of NUV-244 EC50 data

Article Snippet: Lentiviral constructs for PNPLA3 expression , Sirion Biotech , custom.

Techniques: Dispersion

Journal: iScience

Article Title: Identification of NUV-244 as a PNPLA3 I148M degrading small molecule

doi: 10.1016/j.isci.2025.112384

Figure Lengend Snippet: NUV-244 profiling (A) 1.8 μM NUV-244 reduces PNPLA3-tagRFP from the surface of lipid droplets after 24h incubation (white arrows). Hoechst in blue, lipid droplets in yellow (upper panel). Hoechst in blue, PNPLA3-tagRFP in red (lower panel). Scale bar ∼10μm. Representative images shown from n > 3 independent experiments. (B) Dose-response curves for NUV-244 on Huh7 cells either expressing PNPLA3 I148M-tagRFP (black curve) or PNPLA3-tagRFP (red curve). Dose-response with n > 3 per concentration. Curves are normalized to DMSO treated controls (0) and parental Huh7 cells (not expressing PNPLA3, -100). Differences in effect sizes are mainly due to higher total expression levels of PNPLA3 I148M-tagRFP as compared to PNPLA3-tagRFP. Representative data shown from n = 3 independent experiments. (C and D) 1.8 μM NUV-244 reduces untagged PNPLA3 I148M (c.) and PNPLA3 (d.) from the surface of lipid droplets after 24h incubation (white arrows). Hoechst in blue, lipid droplets in yellow (upper panel). Hoechst in blue, Immunofluorescence against PNPLA3 I148M (c.) or PNPLA3 (d.) in red (lower panel). Scale bar ∼10μm. Representative images shown from n > 3 independent experiments. (E) Dose-response curves for NUV-244 on anti-PNPLA3 immunofluorescence quantification of PNPLA3 abundance on lipid droplets in Huh7 cells either expressing PNPLA3 I148M (black curve) or PNPLA3 (red curve). Dose-response with n > 3 per concentration. Curves are normalized to DMSO treated PNPLA3 I148M expressing or PNPLA3 expressing controls (0) and parental Huh7 cells (not expressing PNPLA3, -100). Differences in effect sizes are mainly due to higher total expression levels of PNPLA3 I148M as compared to PNPLA3. Representative data shown from n = 3 independent experiments. (F) Dose-response curves for NUV-244 on Huh7 cells either expressing PNPLA3 I148M-HiBiT (black curve) or PNPLA3-HiBiT (red curve). HiBiT quantification of total protein levels. Dose-response with n > 3 per concentration. Curves are normalized to DMSO treated PNPLA3 I148M-HiBiT expressing or PNPLA3-HiBiT expressing controls (0) and parental Huh7 cells (not expressing PNPLA3, -100). Differences in effect sizes are mainly due to higher total expression levels of PNPLA3 I148M-HiBiT as compared to PNPLA3-HiBiT. Representative data shown from n = 3 independent experiments.

Article Snippet: Lentiviral constructs for PNPLA3 expression , Sirion Biotech , custom.

Techniques: Incubation, Expressing, Concentration Assay, Immunofluorescence

Journal: iScience

Article Title: Identification of NUV-244 as a PNPLA3 I148M degrading small molecule

doi: 10.1016/j.isci.2025.112384

Figure Lengend Snippet: NUV-244 leads to increase in lipid droplet numbers (A) In contrast to Huh7 parental cells (lower picture), lipid droplets are clustered around the nucleus in cells expressing PNPLA3 I148M-tagRFP (white arrows in upper picture). Hoechst in blue, PNPLA3 I148M-tagRFP in red, lipid droplets in yellow. Scale bar ∼10μm. Representative images shown from n > 3 independent experiments. (B) HTS hits at retest (5μM concentration) stage showing scatterplot with the relationship of HTS hits leading to PNPLA3 I148M-tagRFP reduction on lipid droplets (X axis) and lipid droplet count in the same cells. Greener color (color coded for Z score on HiBiT level reduction: blue low/no activity, green higher activity) indicates higher activity in the Huh7 PNPLA3 I148M-HiBiT assay, showing that small molecule-induced reduction in PNPLA3 I148M levels is frequently linked to a decrease in total PNPLA3 I148M protein. (C) EC50 values of NUV-244 (resynthesis) for reduction of PNPLA3 I148M-tagRFP (black curve) on lipid droplets and the normalization of lipid droplet morphology (reduction in lipid droplet aggregates, red curve). Dose-response curves normalized to DMSO treated Huh7 (PNPLA3 I148M-tagRFP) (0) and parental Huh7 (−100) controls. Dose-response with n > 3 per concentration. Representative data shown from n = 3 independent experiments.

Article Snippet: Lentiviral constructs for PNPLA3 expression , Sirion Biotech , custom.

Techniques: Expressing, Concentration Assay, Activity Assay

Journal: iScience

Article Title: Identification of NUV-244 as a PNPLA3 I148M degrading small molecule

doi: 10.1016/j.isci.2025.112384

Figure Lengend Snippet: NUV-244 mediated degradation of PNPLA3 I148M-tagRFP occurs through the 26S proteasome and involves the E3 ligase BFAR (A) Huh7 cells expressing PNPLA3 I148M-tagRFP were treated with DMSO, 1μM or 5μM NUV-244 and co-treated either with DMSO or 1μM Bortezomib or 5μM MLN7243 and subjected to time-lapse imaging for 20h with one image taken every 15 min. Pictures are stills from timelapse quantified in b. at timepoint t = 0h and t = 10h. Scale bar ∼10μm. Representative images shown from n = 2 independent experiments. (B) Quantification of PNPLA3 I148M-tagRFP on lipid droplets shows rapid decrease in PNPLA3 I148M-tagRFP intensity on lipid droplets following treatment of 1μM or 5μM NUV-244 in cells (black and gray curves) and rescue of degradation after 1μM Bortezomib (red curves) or 5μM MLN7243 (blue curves) co-treatment but not with 5μM MLN4929 (green curves). All values are background corrected tagRFP intensity measurements on lipid droplets normalized to DMSO control and t = 0. n = 9 per condition. Mean and standard deviation (error bars); two independent experiments with comparable results. (C) Huh7 cells expressing PNPLA3 I148M-tagRFP were treated with non-targeting control (left) or BFAR siRNA (right) for 48h, treated with DMSO, 1μM or 5μM NUV-244 and subjected to time-lapse imaging for 20h with one image taken every 15 min. Pictures are stills from 1μM NUV-244 treated cells from timelapse quantified in d. at timepoint t = 0h and t = 10h. Scale bar ∼10μm. Representative images shown from n = 2 independent experiments. (D) Quantification of PNPLA3 I148M-tagRFP on lipid droplets shows rapid decrease in PNPLA3 I148M-tagRFP intensity on lipid droplets following treatment of 1μM or 5μM NUV-244 in cells treated with non-targeting control siRNA (black and gray curves) and a rescue of degradation after BFAR knockdown (red curves). tagRFP intensity measurements on lipid droplets normalized to DMSO control and t = 0. n = 9 per condition. Mean and standard deviation (error bars); two independent experiments with comparable results.

Article Snippet: Lentiviral constructs for PNPLA3 expression , Sirion Biotech , custom.

Techniques: Expressing, Imaging, Control, Standard Deviation, Knockdown

Journal: iScience

Article Title: Identification of NUV-244 as a PNPLA3 I148M degrading small molecule

doi: 10.1016/j.isci.2025.112384

Figure Lengend Snippet: NUV-244 does not affect proliferative fitness in liver cells and restores proliferative fitness in PNPLA3 I148M expressing cells (A) Growth curves of Huh7, Hep3B and LX-2 cells were measured in Incucyte. NUV-244 at 5μM (red curves) shows no signs of toxicity compared to DMSO treated controls (black curves). Similar data was obtained with 1μM NUV-244 (data not shown). n = 16 per condition. Mean and standard deviation (error bars); two independent experiments with comparable results. (B) Expression of PNPLA3 I148M in Huh7 cells reduces proliferative fitness (black curve) compared to non-expressing parental control cells (red curve). Treatment with 5μM NUV-244 leads to restoration of cellular proliferative fitness (blue curve; similar data with 1μM NUV-244). n = 16 per condition. Mean and standard deviation (error bars); two independent experiments with comparable results. Highly significant (adjusted p value < 0.0001) differences in proliferation can be seen in Huh7 expressing PNPLA3 I148M treated with NUV-244 compared to DMSO control from 100h incubation onward (two-way ANOVA, not shown).

Article Snippet: Lentiviral constructs for PNPLA3 expression , Sirion Biotech , custom.

Techniques: Expressing, Standard Deviation, Control, Incubation

Journal: iScience

Article Title: Identification of NUV-244 as a PNPLA3 I148M degrading small molecule

doi: 10.1016/j.isci.2025.112384

Figure Lengend Snippet:

Article Snippet: Lentiviral constructs for PNPLA3 expression , Sirion Biotech , custom.

Techniques: Recombinant, Construct, Expressing, Software

Journal: Nature Communications

Article Title: Loss of heterozygosity of essential genes represents a widespread class of potential cancer vulnerabilities

doi: 10.1038/s41467-020-16399-y

Figure Lengend Snippet: a Schematic indicating allele-specific CRISPR approach. “Preexisting genome” represents individuals heterozygous for a germline SNP in a S. pyogenes Cas9 protospacer adjacent motif (PAM) site. A “G” allele (blue) in the PAM retains Cas9 activity at the target site, making this allele CRISPR-sensitive (S). An allele other than “G,” represented by “X” (red) abrogates Cas9 activity at the target site, making this allele CRISPR-resistant (R). Expression of an allele-specific (AS) CRISPR sgRNA targeting the polymorphic PAM site leads to specific inactivation of the S allele. b Schematic of PRIM1 SNP rs2277339 locus showing target sites for positive control, non-allele specific (NA) sgRNA and experimental, allele-specific (AS) sgRNA. Alleles appear in bold. c Crystal structure of PRIM1 gene product shows the amino acid encoded by rs2277339 (teal) lies on the surface of the primase catalytic subunit (gray) near a potentially small-molecule accessible location. d Immunoblot of PRIM1 protein levels in indicated patient-derived cell lines expressing LacZ, PRIM1 NA, or PRIM1 AS sgRNA ( n = 1 biological replicate). e Representative growth curves of indicated patient-derived cell lines expressing LacZ (black), PRIM1 NA (red), or PRIM1 AS (blue) sgRNA, as measured by CellTiter-Glo luminescence, relative to day of assay plating. n = 5 technical replicates. Data are presented as mean values ± s.d. See Supplementary Fig. for additional biological replicates. f Representative growth curves of indicated isogenic cell lines expressing LacZ (black), PRIM1 NA (red), or PRIM1 AS (blue) sgRNA, as measured by CellTiter-Glo luminescence, relative to day of assay plating. n = 5 technical replicates. Data are presented as mean values ± s.d. See Supplementary Fig. for additional biological replicates. g Disruption of PRIM1 in isogenic hemizygous PRIM1 resistant (PRIM1 R ) or PRIM1 sensitive (PRIM1 S ) cells expressing PRIM1 NA or AS sgRNA. Unaltered alleles (black), alleles with in-frame insertions or deletions (gray), and alleles with frameshift alterations (yellow) were assessed by deep sequencing of PRIM1 four days post-infection with sgRNA. Source data for Fig. 2d–g are provided as a Source Data file.

Article Snippet: Lentiviral expression constructs for non-allele specific shRNA-mediated suppression of PRIM1 were obtained through the Broad Institute of MIT and Harvard Genomic Perturbation Platform ( https://portals.broadinstitute.org/gpp/public/ ).

Techniques: CRISPR, Activity Assay, Expressing, Positive Control, Western Blot, Derivative Assay, Disruption, Sequencing, Infection

Journal: Nature Communications

Article Title: Loss of heterozygosity of essential genes represents a widespread class of potential cancer vulnerabilities

doi: 10.1038/s41467-020-16399-y

Figure Lengend Snippet: a Number of GEMINI variants (vertical axis) plotted against the number of patients per year in the US whose tumors might respond to therapeutics targeting those variants (i.e., have lost the resistant allele from a heterozygous germline; horizontal axis). Bin width = 1000 patients. b Growth of heterozygous (red circles) and hemizygous cells (pink circles) expressing positive control, non-allele specific PRIM1 -targeting shRNAs versus PRIM1 mRNA expression. Cell growth measured by CellTiter-Glo luminescence relative to day 2 post-infection and shGFP ( n = 5 technical replicates). PRIM1 mRNA expression assessed by qRT-PCR ( n = 3 technical replicates). Data are presented as mean values ± s.d. Dashed gray line indicates PRIM1 expression threshold below which substantial decreases in cell viability are observed. c Summary table representing challenges to developing allele-specific small molecules that target GEMINI vulnerabilities and associated analyses to prioritize targets. Source data for Fig. 4b is provided as a Source Data file.

Article Snippet: Lentiviral expression constructs for non-allele specific shRNA-mediated suppression of PRIM1 were obtained through the Broad Institute of MIT and Harvard Genomic Perturbation Platform ( https://portals.broadinstitute.org/gpp/public/ ).

Techniques: Expressing, Positive Control, Infection, Quantitative RT-PCR