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Dr. Son's mRNA Vaccine Design, First Steps II

You must annotate
before you can choose

Part I handed you one sequence. Open the file and there are 1273 letters in a row. Which of them goes into the vaccine? You cannot say - every letter looks the same.
This volume attaches coordinates of meaning to that string. Pfam matches against what is already known; MEME discovers without knowing. IEDB points at what the immune system might see, and ElliPro pulls out the sites that only appear once the protein folds.
You finish with four numbers: 29,620 / 5,671 / 1,266 / 10. The real subject of this volume is why those numbers are not evidence.
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๐Ÿ Python๐Ÿ”– annotation๐Ÿงฑ domains ๐Ÿ” motifs๐Ÿ‘๏ธ epitopes๐Ÿ“ limits of prediction๐Ÿ‡ฌ๐Ÿ‡ง English
Start lesson 1 โ†’

How you'll learn

๐Ÿ—บ๏ธDraw the map first

You cannot choose from a bare string. Annotation attaches coordinates of meaning to a sequence.

๐Ÿ”งOpen the tools up

Pfam, MEME and IEDB differ only on the surface. Inside, all of them slide a window and score it.

๐Ÿ“Ask the unit first

1,266 (stretches) and 10 (clusters) are different units. Compare counts without checking, and the number lies.

๐ŸšงDon't be fooled by a long list

29,620 is how much was scanned, not how much was found. A multiplication is not an achievement.

Full curriculum

01
๋งจ ๋ฌธ์ž์—ด์—์„œ๋Š” ์•„๋ฌด๊ฒƒ๋„ ๊ณ ๋ฅผ ์ˆ˜ ์—†๋‹ค
Step 2 ยท 1273 ๊ธ€์ž์™€ Fig. 4 ์˜ ๋‘˜์งธ ์นธ
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02
๋„๋ฉ”์ธ๊ณผ ๋ชจํ‹ฐํ”„์™€ ์—ํ”ผํ† ํ”„๋Š” ์„œ๋กœ ๋‹ค๋ฅธ ๊ฒƒ์ด๋‹ค
Step 2 ยท ์ž์ฃผ ๋’ค์„ž์ด๋Š” ๋‚ฑ๋ง ์…‹
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03
Pfam โ€” ์ด๋ฏธ ์•Œ๋ ค์ง„ ๊ฒƒ๊ณผ ๋งž์ถฐ ๋ณธ๋‹ค
Step 2 ยท ๋„๋ฉ”์ธ ์ฃผ์„ ยท pfam.xfam.org
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04
MEME โ€” ์‚ฌ์ „์— ์—†๋Š” ๋ฌด๋Šฌ๋ฅผ ์ฐพ์•„๋‚ธ๋‹ค
Step 2 ยท ๋ชจํ‹ฐํ”„ ๋ฐœ๊ฒฌ ยท meme-suite.org
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05
๋„ˆ๋น„๊ฐ€ ๋œป์„ ๋‚˜๋ฅธ๋‹ค โ€” 6~15 ์™€ 20~50
Step 2 ยท ๋ชจํ‹ฐํ”„ ๋„ˆ๋น„ 6~50 ์•„๋ฏธ๋…ธ์‚ฐ
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06
์„ค์ • ํ•˜๋‚˜๊ฐ€ ์งˆ๋ฌธ์„ ๋ฐ”๊พผ๋‹ค โ€” ํ•˜๋‚˜๋งŒ ์ธ๊ฐ€ ์—ฌ๋Ÿฟ ์ธ๊ฐ€
Step 2 ยท zero or one / zero or more
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07
์ฐฝ์„ ๊ตด๋ฆฐ๋‹ค โ€” ์˜ˆ์ธก ๋„๊ตฌ๊ฐ€ ์†์œผ๋กœ ํ•˜๋Š” ์ผ
Step 2 ยท ์†์„ ์›€์ง์ด๋Š” ์ฒซ ๊ฐ•
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08
๋‘ ๊ฐˆ๋ž˜๋ฅผ ๋‹ค ๊ณจ๋ผ์•ผ ํ•˜๋Š” ๊นŒ๋‹ญ
Step 2 ยท MHC class I ๊ณผ CD8 ยท class II ์™€ CD4
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09
27๊ฐœ๋กœ 97% โ€” ๋Œ€๋ฆฝ์œ ์ „์ž๋ฅผ ๊ณ ๋ฅด๋Š” ์…ˆ๋ฒ•
Step 2 ยท IEDB tc1 ยท 9 ์•„๋ฏธ๋…ธ์‚ฐ ยท HLA class I 27๊ฐœ
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10
๊ฐ™์€ ์ ˆ์ฐจ ๋‹ค๋ฅธ ๊ธธ์ด โ€” class II ์ชฝ ์ฐฝ๊ตฌ
Step 2 ยท IEDB tc2 ยท 15 ์•„๋ฏธ๋…ธ์‚ฐ ยท HLA class II 27๊ฐœ
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11
๋‘ ์ˆซ์ž๊ฐ€ ๊ฐ™์€ ๊ณณ์„ ๊ฐ€๋ฆฌํ‚จ๋‹ค โ€” ์ ์ˆ˜์™€ ํผ์„ผํƒ€์ผ
Step 2 ยท NetMHCpan EL score ยท EL percentile rank
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12
๋ฌธํ„ฑ์„ ๋„๊ตฌ๊ฐ€ ์ •ํ•ด ์ค€๋‹ค โ€” ์„ ํ˜• B์„ธํฌ ์˜ˆ์ธก
Step 2 ยท IEDB B cell ยท BepiPred ยท Karplusโ€“Schulz
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13
๊ตฌ์กฐ๊ฐ€ ์žˆ์–ด์•ผ ๋ณด์ด๋Š” ๊ฒƒ โ€” ์ž…์ฒด ์—ํ”ผํ† ํ”„
Step 2 ยท ElliPro ยท PDB 6VXX ยท ์‚ฌ์Šฌ AยทBยทC
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14
์„ ํ˜•์ด ๋Œ€์ ‘๋ฐ›๋Š” ๊ฒƒ์€ ์˜ณ์•„์„œ๊ฐ€ ์•„๋‹ˆ๋‹ค
Step 2 ยท ์„ ํ˜• ๋Œ€ ์ž…์ฒด ยท ์˜ˆ์ธก ๊ฐ€๋Šฅ์„ฑ์˜ ํŽธํ–ฅ
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15
๋„ค ์ž๋ฆฌ ์ˆซ์ž๋ฅผ ์ฒ˜์Œ ๋งŒ๋‚œ๋‹ค โ€” 29,620 ยท 5,671 ยท 1,266 ยท 10
Step 2 ๊ฒฐ๊ณผ ยท ์˜ˆ์ธก๋œ ์—ํ”ผํ† ํ”„ ์ „์ฒด
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16
๊ธด ๋ชฉ๋ก์€ ๊ทผ๊ฑฐ๊ฐ€ ์•„๋‹ˆ๋‹ค
Step 2 ยท ํ•จ์ • ยท ์˜ˆ์ธก๊ณผ ๊ด€์ฐฐ์˜ ๊ฑฐ๋ฆฌ
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17
์•ˆ์ชฝ์— ํŒŒ๋ฌปํžŒ ์ž๋ฆฌ๋Š” ์•„๋ฌด๋ฆฌ ์ข‹์•„๋„ ์†Œ์šฉ์—†๋‹ค
Step 2 ยท ํ‘œ๋ฉด ๋…ธ์ถœ๊ณผ ์„ธํฌ์™ธ ์˜์—ญ
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18
์‚ฌ๋žŒ์ด ์†์œผ๋กœ ํ™•์ธํ•ด ๋‘” ๊ฒƒ์ด ์žˆ๋‹ค๋ฉด ๊ทธ๊ฒƒ๋ถ€ํ„ฐ ์“ด๋‹ค
Step 2 ยท UniProt Feature Viewer ยท GFF ๋‚ด๋ ค๋ฐ›๊ธฐ
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19
์ฃผ์„์„ ์ฒ˜์Œ๋ถ€ํ„ฐ ๋๊นŒ์ง€ ๋‹ฌ์•„ ๋ณธ๋‹ค
Step 2 ์ข…ํ•ฉ ยท ํ‘œ ๋„ท์„ ์ง์ ‘ ๋งŒ๋“ ๋‹ค
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20
์ฃผ์„์„ ๋งˆ์นœ ์ž๋ฆฌ์—์„œ ๋งํ•  ์ˆ˜ ์žˆ๋Š” ๊ฒƒ
Step 2 ์ •๋ฆฌ ยท ๊ทธ๋ฆฌ๊ณ  ๋งํ•  ์ˆ˜ ์—†๋Š” ๊ฒƒ
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