Research
Publications
These papers exist because we built the platform and needed the work to live somewhere permanent. The main preprint documents the full computational pipeline — from RNA-seq through the vanillylamine discovery to synthesis-ready DNA. The cross-species paper is the follow-on: the same pathway engineered into 17 other plant families. Nine more papers covering the full Iyengar Flavor Index platform are in preparation. All capsaicin DOIs go live on October 31, 2026 at 8:08 PM PT.
Author: Priyanca Ford · ORCID: 0000-0003-0395-1752 · Scoville Splice LLC
Primary Research Article · Paper 1
Computational Design of Ten Superhot Capsicum Cultivars: Vanillylamine Precursor Supply, Not Pun1/AT3, Controls the Scoville Ceiling
Integrated Transcriptomic, Metabolic, and Synthetic Biology Pipeline for Engineered Capsaicinoid Accumulation
An integrated computational pipeline combining developmental RNA-seq (Salmon 2.8.0), PyDESeq2 differential expression (4,365 significant genes, 28,074 tested), constraint-based metabolic flux modeling (COBRApy 0.32.1, 37 metabolites, 44 reactions), ESM2 protein language model mutagenesis (650M parameters), and CRISPR-SpCas9 construct design produces ten engineered Capsicum cultivar specifications spanning 866,611–2,956,310 SHU. Analysis of 4,365 differentially expressed genes across a C. chinense developmental time series identifies vanillylamine precursor supply — not Pun1/AT3 acyltransferase activity — as the principal bottleneck controlling the capsaicinoid ceiling in existing high-SHU germplasm.
Cross-Species Research · Paper 2
Capsaicin Without Capsicum: Cross-Species Capsaicinoid Pathway Engineering Across 17 Plant Species
Engineering the capsaicinoid biosynthesis pathway beyond Capsicum into 17 non-pepper plant species. Demonstrates that 5 of 7 platform modules transfer unchanged across organisms — codon re-optimization, tissue-specific promoters, and species-validated transformation protocols are the only organism-specific work required. Provides a complete cross-species engineering handbook with 47 codon-optimized construct sequences deposited in NCBI GenBank.
Iyengar Flavor Index & Platform Papers · Papers 3–11
Nine papers covering the full IFI flavor engineering platform — receptor-axis construct specifications for sweetness, umami, bitterness, kokumi, astringency, cool-burn, TRPA1 pungency, and the overarching platform framework. DOIs registered; PDFs in preparation.
Sweet Heat: Brazzein Co-Expression and Capsaicinoid Dual Engineering via IFI-H × IFI-S Receptor Axes
The Iyengar Flavor Index: A Receptor-Normalized Seven-Axis Quantitative Scale for Full-Spectrum Chemosensory Engineering
Umami-Enriched Capsicum via GAD1/4 Knockout and GDH1 Overexpression: IFI-U Construct Specifications
Zero-Bitter Brassica: TGG1/TGG2 Myrosinase Knockout for IFI-B Construct Specifications
Kokumi Mouthfulness Engineering via GCS1 Overexpression for γ-Glutamyl Peptide Accumulation: IFI-K Construct Specifications
Astringency Reduction via ANR Knockout in Grapes and Sorghum: IFI-A Construct Specifications
Cool-Burn: Five-Gene Menthol Pathway Engineering with IFI-C ~85,000 via TRPM8/TRPV1 Co-Modulation
TRPA1 Spectrum Engineering: CrCCR1 Cinnamaldehyde Pathway for IFI-P Pungency Construct Specifications
The Scoville Splice Capsaicin Design Platform: An Integrated Computational and Synthetic Biology Framework for Full-Spectrum Flavor Engineering
Cultivar Specification Papers
Ten individual cultivar specifications, ranked #1–#10 by computational SHU target.
#1 of 10
2,956,310
SHU
Maximum computational capsaicinoid target using complete pathway rewiring with ESM2-optimized Pun1 triple mutant (S39L+L345G+C175S). At 200 mg/g dry weight (20.0% DW), this design represents the computational ceiling for the four-gene intervention stack.
#2 of 10
2,802,656
SHU
Near-ceiling Tier C computational design via full four-intervention capsaicinoid pathway rewiring. Complete four-gene stack with ESM2-optimized Pun1 double mutant achieves 95% of the theoretical maximum.
#3 of 10
2,458,470
SHU
Triple-gene precursor stack plus Pun1 double mutant (S39L+L345G) achieves 2.46M SHU via enzyme-level and precursor-level synergy in C. chinense placental tissue.
#4 of 10
2,267,939
SHU
Maximum Tier B phenylpropanoid stack achieves 2.27M SHU via coordinated three-gene intervention in C. chinense. Exceeds the disputed Pepper X figure on a per-capsaicinoid efficiency basis.
#5 of 10
2,065,115
SHU
Targets 2.07M SHU, crossing the verified Carolina Reaper Guinness World Record (1,641,183 SHU) by 26%. At 145 mg/g dry weight (14.5% DW), capsaicinoid concentration is 37% above the measured physical precedent ceiling.
#6 of 10
1,849,999
SHU
Dual precursor amplification via PAL and COMT overexpression enables sub-2M SHU above the physical dry-weight ceiling in engineered C. chinense. First design requiring speculative transcriptional reprogramming.
#7 of 10
1,622,590
SHU
Borderline-feasibility Tier A design targeting certified world-record-class SHU via a single CRISPR intervention beyond the measured capsaicinoid dry-weight ceiling. Approaches the verified Carolina Reaper record.
#8 of 10
1,382,889
SHU
Near-maximum achievable capsaicinoid concentration via single-gene peroxidase knockout at exactly 100 mg/g dry weight (10.0% DW). Approaches the measured physical ceiling while remaining within documented precedent.
#9 of 10
1,130,896
SHU
Targets 1.13M SHU — decisively exceeding the Bhut jolokia (1,041,427 SHU) and well within the single-gene peroxidase knockout feasibility envelope. At 82 mg/g dry weight (8.2% DW).
#10 of 10
866,611
SHU
Entry-level cultivar targeting 866,611 SHU via a single CRISPR-SpCas9 intervention: knockout of the competing peroxidase gene that diverts vanillylamine precursors away from the capsaicinoid pathway. At 64 mg/g dry weight (6.4% DW).
Data & Sequence Repositories
bioRxiv · Preprints
- Computational Design of Ten Superhot Capsicum Cultivars ↗
- Beyond Peppers: Cross-Species Capsaicinoid Engineering ↗
Submitted · Live within 24–48 hrs of Oct 31, 2026
Harvard Dataverse
Full collection (12 papers) — 10.7910/DVN/BO05NO ↗- ✓Computational Design of Ten Cultivars · 10.7910/DVN/BO05NO
- ⏳Beyond Peppers · 10.7910/DVN/GO8MRF
- ⏳SS-0087 Verbal Warning · 10.7910/DVN/KDAKPO
- ⏳SS-0088 Kill Switch · 10.7910/DVN/ELNYA2
- ⏳SS-0089 Hostile Agent · 10.7910/DVN/H4HVBS
- ⏳SS-0090 Containment Breach · 10.7910/DVN/S5RTJW
- ⏳SS-0091 System Shock · 10.7910/DVN/BXZZ8P
- ⏳SS-0092 Venom Protocol · 10.7910/DVN/B59QT4
- ⏳SS-0093 Level 4 Pathogen · 10.7910/DVN/HYZ2OA
- ⏳SS-0094 Collateral Damage · 10.7910/DVN/DGFAFU
- ⏳SS-0095 Extinction Protocol · 10.7910/DVN/EM8CWO
- ⏳SS-0096 D.N.R. · 10.7910/DVN/1VNFQ8
NCBI
- BioProject PRJNA1554082 — Capsicum Transcriptomics & Construct Sequences ↗
- BioSample batch SAMN64571838–SAMN64571847 (10 cultivars) ↗
- GenBank submission SUB16596390 · pending
NCBI GenBank · 40 sequences
CRISPR-SpCas9 Guide Sequences (QB169135–QB169174)
Embargoed pending patent · Accesses first sequence in range
Figshare
Scoville Splice — Full Dataset (16 files)
10.6084/m9.figshare.34875789 ↗
Open Science Framework
Scoville Splice — Full Project (16 files)
10.17605/OSF.IO/756YJ ↗
NCBI GenBank · Pending Accessions
- SUB165963906 Capsicum synthetic construct components
- SUB1659634847 cross-species codon-optimized constructs
- SUB16596317Pun1 triple mutant CDS (SS-PUN1-TM)
- SUB16603688Brazzein + capsaicinoid constructs (Paper 3)
- SUB16603765GAD1/4 KO + GDH1 OE constructs (Paper 5)
- SUB16603811TGG1/TGG2 KO constructs (Paper 6)
- SUB16603848GCS1 OE constructs (Paper 7)
- SUB16603898ANR KO constructs (Paper 8)
- SUB166039475-gene menthol pathway constructs (Paper 9)
- SUB16603959CrCCR1 cinnamaldehyde constructs (Paper 10)
How to Cite
Questions about the research? scovillesplice@gmail.com