FLASH assembly of TALENs for high-throughput genome editing.
Reyon D, Tsai SQ, Khayter C, Foden JA, Sander JD, Joung JK
Nat Biotechnol. 2012 Apr 8. doi: 10.1038/nbt.2170.
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PubMed
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Article
The Joung Lab used their high-throughput FLASH method to construct one large series of engineered TALENs targeted to 48 sites in the EGFP reporter gene and another large series targeted to human genes involved in cancer and/or epigenetic regulation.
Engineered TALENs for the EGFP Reporter Gene
The table below lists plasmids encoding these 48 pairs of TALENs that were shown to be active at their target sites on an integrated EGFP reporter gene in cultured U2OS cells.
| EGFP TALEN Plasmid | Plasmid Name | Target Half-Site |
|---|---|---|
| eGFP-TALEN-01-Left | TAL2000 | TCGCCACCAT |
| eGFP-TALEN-01-Right | TAL2001 | TGAACAGCTC |
| eGFP-TALEN-02-Left | TAL2002 | TGGTGCCCAT |
| eGFP-TALEN-02-Right | TAL2003 | TTACGTCGCC |
| eGFP-TALEN-03-Left | TAL2004 | TCTGCACCAC |
| eGFP-TALEN-03-Right | TAL2005 | TGGGCCAGGG |
| eGFP-TALEN-04-Left | TAL2006 | TGGAGTACAA |
| eGFP-TALEN-04-Right | TAL2007 | TGATATAGAC |
| eGFP-TALEN-05-Left | TAL2008 | TTCAGCGTGTC |
| eGFP-TALEN-05-Right | TAL2009 | TAGGTGGCATC |
| eGFP-TALEN-06-Left | TAL2010 | TGCCACCTACG |
| eGFP-TALEN-06-Right | TAL2011 | TGCAGATGAAC |
| eGFP-TALEN-07-Left | TAL2012 | TGGCCCACCCT |
| eGFP-TALEN-07-Right | TAL2013 | TGCACGCCGTA |
| eGFP-TALEN-08-Left | TAL2014 | TTCAAGATCCG |
| eGFP-TALEN-08-Right | TAL2015 | TGCACGCTGCC |
| eGFP-TALEN-09-Left | TAL2016 | TAGAGGATCCAC |
| eGFP-TALEN-09-Right | TAL2017 | TCGCCCTTGCTC |
| eGFP-TALEN-10-Left | TAL2018 | TCCGGCGAGGGC |
| eGFP-TALEN-10-Right | TAL2019 | TCAGCTTGCCGT |
| eGFP-TALEN-11-Left | TAL2020 | TGACCTACGGCG |
| eGFP-TALEN-11-Right | TAL2021 | TGGTCGGGGTAG |
| eGFP-TALEN-12-Left | TAL2022 | TCCGCCACAACA |
| eGFP-TALEN-12-Right | TAL2023 | TCGGCGAGCTGC |
| eGFP-TALEN-13-Left | TAL2024 | TCCTGGTCGAGCT |
| eGFP-TALEN-13-Right | TAL2025 | TGAACTTGTGGCC |
| eGFP-TALEN-14-Left | TAL2026 | TCAGCCGCTACCC |
| eGFP-TALEN-14-Right | TAL2027 | TGAAGAAGTCGTG |
| eGFP-TALEN-15-Left | TAL2028 | TCTTCAAGTCCGC |
| eGFP-TALEN-15-Right | TAL2029 | TGCGCTCCTGGAC |
| eGFP-TALEN-16-Left | TAL2030 | TCAAGGAGGACGG |
| eGFP-TALEN-16-Right | TAL2031 | TGTACTCCAGCTT |
| eGFP-TALEN-17-Left | TAL2032 | TCAAGGTGAACTT |
| eGFP-TALEN-17-Right | TAL2033 | TGCCGTCCTCGAT |
| eGFP-TALEN-18-Left | TAL2034 | TCCACCGGTCGCCA |
| eGFP-TALEN-18-Right | TAL2035 | TGAACAGCTCCTCG |
| eGFP-TALEN-19-Left | TAL2036 | TTCAGCGTGTCCGG |
| eGFP-TALEN-19-Right | TAL2037 | TTGCCGTAGGTGGC |
| eGFP-TALEN-20-Left | TAL2038 | TTCAGCCGCTACCC |
| eGFP-TALEN-20-Right | TAL2039 | TTGAAGAAGTCGTG |
| eGFP-TALEN-21-Left | TAL2040 | TACAACAGCCACAA |
| eGFP-TALEN-21-Right | TAL2041 | TTCTTCTGCTTGTC |
| eGFP-TALEN-22-Left | TAL2042 | TGGTGCCCATCCTGG |
| eGFP-TALEN-22-Right | TAL2043 | TTGTGGCCGTTTACG |
| eGFP-TALEN-23-Left | TAL2044 | TGCCCGAAGGCTACG |
| eGFP-TALEN-23-Right | TAL2045 | TCGTCCTTGAAGAAG |
| eGFP-TALEN-24-Left | TAL2046 | TGAACCGCATCGAGC |
| eGFP-TALEN-24-Right | TAL2047 | TTGCCGTCCTCCTTG |
| eGFP-TALEN-25-Left | TAL2048 | TCAAGATCCGCCACA |
| eGFP-TALEN-25-Right | TAL2049 | TCGGCGAGCTGCACG |
| eGFP-TALEN-26-Left | TAL2050 | TGTTCACCGGGGTGGT |
| eGFP-TALEN-26-Right | TAL2051 | TTACGTCGCCGTCCAG |
| eGFP-TALEN-27-Left | TAL2052 | TGCCCATCCTGGTCGA |
| eGFP-TALEN-27-Right | TAL2053 | TGAACTTGTGGCCGTT |
| eGFP-TALEN-28-Left | TAL2054 | TGTCCGGCGAGGGCGA |
| eGFP-TALEN-28-Right | TAL2055 | TCAGGGTCAGCTTGCC |
| eGFP-TALEN-29-Left | TAL2056 | TCATCTGCACCACCGG |
| eGFP-TALEN-29-Right | TAL2057 | TCACGAGGGTGGGCCA |
| eGFP-TALEN-30-Left | TAL2058 | TCTATATCATGGCCGA |
| eGFP-TALEN-30-Right | TAL2059 | TGAAGTTCACCTTGAT |
| eGFP-TALEN-31-Left | TAL2060 | TACGGCGTGCAGTGCTT |
| eGFP-TALEN-31-Right | TAL2061 | TCGTGCTGCTTCATGTG |
| eGFP-TALEN-32-Left | TAL2086 | TACCCCGACCACATGAA |
| eGFP-TALEN-32-Right | TAL2087 | TCGGGCATGGCGGACTT |
| eGFP-TALEN-33-Left | TAL2062 | TCCGCCATGCCCGAAGG |
| eGFP-TALEN-33-Right | TAL2063 | TCCTTGAAGAAGATGGT |
| eGFP-TALEN-34-Left | TAL2090 | TTCAAGGAGGACGGCAA |
| eGFP-TALEN-34-Right | TAL2091 | TTGTAGTTGTACTCCAG |
| eGFP-TALEN-35-Left | TAL2092 | TACAACTACAACAGCCA |
| eGFP-TALEN-35-Right | TAL2093 | TTCTTCTGCTTGTCGGC |
| eGFP-TALEN-36-Left | TAL2094 | TCACCGGGGTGGTGCCCA |
| eGFP-TALEN-36-Right | TAL2095 | TGGCCGTTTACGTCGCCG |
| eGFP-TALEN-37-Left | TAL2096 | TAAACGGCCACAAGTTCA |
| eGFP-TALEN-37-Right | TAL2097 | TAGGTGGCATCGCCCTCG |
| eGFP-TALEN-38-Left | TAL2098 | TGCAGTGCTTCAGCCGCT |
| eGFP-TALEN-38-Right | TAL2099 | TTGAAGAAGTCGTGCTGC |
| eGFP-TALEN-39-Left | TAL2064 | TCAGCGTGTCCGGCGAGGG |
| eGFP-TALEN-39-Right | TAL2065 | TCAGGGTCAGCTTGCCGTA |
| eGFP-TALEN-40-Left | TAL2066 | TGAAGTTCATCTGCACCAC |
| eGFP-TALEN-40-Right | TAL2067 | TCACGAGGGTGGGCCAGGG |
| eGFP-TALEN-41-Left | TAL2068 | TCGTGACCACCCTGACCTA |
| eGFP-TALEN-41-Right | TAL2069 | TGTGGTCGGGGTAGCGGCT |
| eGFP-TALEN-42-Left | TAL2070 | TCATGGCCGACAAGCAGAA |
| eGFP-TALEN-42-Right | TAL2071 | TGTGGCGGATCTTGAAGTT |
| eGFP-TALEN-43-Left | TAL2072 | TTCACCGGGGTGGTGCCCAT |
| eGFP-TALEN-43-Right | TAL2073 | TTGTGGCCGTTTACGTCGCC |
| eGFP-TALEN-44-Left | TAL2074 | TGCACCACCGGCAAGCTGCC |
| eGFP-TALEN-44-Right | TAL2075 | TAGGTCAGGGTGGTCACGAG |
| eGFP-TALEN-45-Left | TAL2076 | TTCAAGTCCGCCATGCCCGA |
| eGFP-TALEN-45-Right | TAL2077 | TCCTTGAAGAAGATGGTGCG |
| eGFP-TALEN-46-Left | TAL2078 | TATATCATGGCCGACAAGCA |
| eGFP-TALEN-46-Right | TAL2079 | TGGCGGATCTTGAAGTTCAC |
| eGFP-TALEN-47-Left | TAL2080 | TTCATCTGCACCACCGGCAAG |
| eGFP-TALEN-47-Right | TAL2081 | TCAGGGTGGTCACGAGGGTGG |
| eGFP-TALEN-48-Left | TAL2084 | TGAAGTTCGAGGGCGACACCC |
| eGFP-TALEN-48-Right | TAL2085 | TTGAAGTCGATGCCCTTCAGC |
Engineered TALENs for Endogenous Human Gene Targets
The table below lists 84 pairs of these TALENs that were shown to be active at their target sites in cultured U2OS cells.
| Target Gene Name | TALENs | Full Target Site (5' to 3'; half-sites in CAPS, spacer in lowercase) |
|---|---|---|
| ABL1 | TAL2380 & TAL2381 | TACCTATTATTACTTTATggggcagcagcctggaaAAGTACTTGGGGACCAA |
| AKT2 | TAL2382 & TAL2383 | TGTGTCTTGGGATGAGTGggtcagtgttctggtgCTCACAGGATGGCTGGCA |
| ALK | TAL2292 & TAL2293 | TCCTGTGGCTCCTGCCGCtgctgctttccacggcAGCTGTGGGCTCCGGGA |
| APC | TAL2294 & TAL2295 | TATGTACGCCTCCCTGGGctcgggtccggtcgccCCTTTGCCCGCTTCTGTA |
| ATM | TAL2402 & TAL2403 | TGAATTGGGATGCTGTTTttaggtattctattcaaaTTTATTTTACTGTCTTTA |
| AXIN2 | TAL2200 & TAL2201 | TCCCTCACCATGAGTAGCgctatgttggtgacttGCCTCCCGGACCCCAGCA |
| BAX | TAL2296 & TAL2297 | TGTGCGATCTCCAAGCACtgaggggcagaaactcCCGGATCGGGCGCTGCCA |
| BCL6 | TAL2412 & TAL2413 | TTTTCAAGTGAAGACAAAatggcctcgccggctgACAGCTGTATCCAGTTCA |
| BMPR1A | TAL2260 & TAL2261 | TACAATTGAACAATGCCTcagctatacatttacatCAGATTATTGGGAGCCTA |
| BRCA1 | TAL2384 & TAL2385 | TCCGAAGCTGACAGATGGgtattctttgacggggGGTAGGGGCGGAACCTGA |
| BRCA2 | TAL2302 & TAL2303 | TTAGACTTAGGTAAGTAAtgcaatatggtagactGGGGAGAACTACAAACTA |
| CBX3 | TAL2358 & TAL2359 | TCTGCAATAAAAAATGGCctccaacaaaactacaTTGGTAAGTTAATGAAAA |
| CBX8 | TAL2360 & TAL2361 | TGGAGCTTTCAGCGGTGGgggagcgggtgttcgcgGCCGAAGCCCTCCTGAA |
| CCND1 | TAL2304 & TAL2473 | TGGAACACCAGCTCCTGTgctgcgaagtggaaaccatCCGCCGCGCGTACCCCGA |
| CDC73 | TAL2202 & TAL2203 | TGCTTAGCGTCCTGCGACagtacaacatccagaaGAAGGAGATTGTGGTGAA |
| CDK4 | TAL2404 & TAL2405 | TCCCTTGATCTGAGAAtggctacctctcgataTGAGCCAGTGGCTGAAA |
| CHD4 | TAL2236 & TAL2237 | TGGCGTCGGGCCTGGGCtccccgtccccctgctcGGCGGGCAGTGAGGAGGA |
| CHD7 | TAL2238 & TAL2239 | TGTGTTGGAAGAAGATGGcagatccaggaatgatGAGTCTTTTTGGCGAGGA |
| CTNNB1 | TAL2262 & TAL2263 | TCCAGCGTGGACAATGGctactcaaggtttgtgTCATTAAATCTTTAGTTA |
| CYLD | TAL2386 & TAL2387 | TAATATCACAATGAGTTCaggcttatggagccaagaAAAAGTCACTTCACCCTA |
| DDB2 | TAL2282 & TAL2283 | TCACACGGAGGACGCGatggctcccaagaaacGCCCAGAAACCCAGAAGA |
| ERCC2 | TAL2306 & TAL2307 | TCCGGCCGGCGCCATGAagtgagaagggggctgGGGGTCGCGCTCGCTA |
| EWSR1 | TAL2252 & TAL2253 | TGGCGTCCACGGGTGAGTatggtggaactgcggtcGCGCCGGCGGTAGCCGGA |
| EXT1 | TAL2414 & TAL2415 | TGACCCAGGCAGGACACAtgcaggccaaaaaacgcTATTTCATCCTGCTCTCA |
| EXT2 | TAL2204 & TAL2205 | TTCCTCCCAGGGGGATGTcctgcgcctcagggtcCGGTGGTGGCCTGCGGCA |
| EZH2 | TAL2362 & TAL2363 | TGCTTTTAGAATAATCATgggccagactgggaagAAATCTGAGAAGGGACCA |
| FANCA | TAL2416 & TAL2417 | TAGGCGCCAAGGCCATGTccgactcgtgggtcccGAACTCCGCCTCGGGCCA |
| FANCC | TAL2254 & TAL2255 | TGAAGGGACATCACCTTTtcgctttttccaagatgGCTCAAGATTCAGTAGA |
| FANCF | TAL2314 & TAL2315 | TTCGCGCACCTCATGGaatcccttctgcagcaCCTGGATCGCTTTTCCGA |
| FANCG | TAL2288 & TAL2289 | TCGGCCACCATGTCCCgccagaccacctctgtGGGCTCCAGCTGCCTGGA |
| FES | TAL2390 & TAL2391 | TCCCCAGAACAGCACTATgggcttctcttccgagctGTGCAGCCCCCAGGGCCA |
| FGFR1 | TAL2316 & TAL2317 | TCTGCTCCCCACCGAGGAcctctgcatgcaggcaTGAATCCCAGGAGCCTA |
| FH | TAL2266 & TAL2267 | TGTACCGAGCACTTCGGCtcctcgcgcgctcgcgtCCCCTCGTGCGGGCTCCA |
| FLCN | TAL2206 & TAL2207 | TCTCCAAGGCACCATGAAtgccatcgtggctctctgCCACTTCTGCGAGCTCCA |
| FLT4 | TAL2320 & TAL2321 | TGCAGCGGGGCGCCGCGCtgtgcctgcgactgtggctCTGCCTGGGACTCCTGGA |
| FOXO1 | TAL2322 & TAL2323 | TCACCATGGCCGAGGCGcctcaggtggtggagaTCGACCCGGACTTCGA |
| FOXO3 | TAL2268 & TAL2269 | TCTCCGCTCGAAGTGGAGctggacccggagttcgagCCCCAGAGCCGTCCGCGA |
| GLI1 | TAL2270 & TAL2271 | TCCTCTGAGACGCCATGTtcaactcgatgaccccACCACCAATCAGTAGCTA |
| HDAC1 | TAL2364 & TAL2365 | TGGCGCAGACGCAGGGCacccggaggaaagtctgTTACTACTACGACGGTGA |
| HDAC2 | TAL2430 & TAL2431 | TGCGCTCACCTCCCTGCGgcctcctgaggtggtttgGTGGCCCCCTCCTCGCGA |
| HDAC6 | TAL2400 & TAL2401 | TCCTCAACTATGACCTCAaccggccaggattccaCCACAACCAGGCAGCGAA |
| HMGA2 | TAL2450 & TAL2451 | TGAGCGCACGCGGTGAGGgcgcggggcagccgtcCACTTCAGCCCAGGGACA |
| HOXA13 | TAL2208 & TAL2209 | TCCGTGCTCCTCCACCCCcgctggatcgagcccacCGTCATGTTTCTCTACGA |
| HOXA9 | TAL2444 & TAL2445 | TGGGCACGGTGATGGCcaccactggggccctgGGCAACTACTACGTGGA |
| HOXC13 | TAL2286 & TAL2287 | TCCAGCAGATCATGTCATgacgacttcgctgctcctGCATCCACGCTGGCCGGA |
| JAK2 | TAL2406 & TAL2407 | TCTGAAAAAGACTCTGCAtgggaatggcctgcctTACGATGACAGAAATGGA |
| KRAS | TAL2272 & TAL2273 | TGAAAATGACTGAATATAaacttgtggtagttggaGCTGGTGGCGTAGGCAA |
| MAP2K4 | TAL2324 & TAL2325 | TAGGGTCCCCGGCGCCAGgccacccggccgtcagCAGCATGCAGGGTAAGGA |
| MDM2 | TAL2274 & TAL2275 | TCCAAGCGCGAAAACCCCggatggtgaggagcaggTACTGGCCCGGCAGCGA |
| MET | TAL2326 & TAL2327 | TTATTATTACATGGCTTTgccttactgaggcttcATCTTGTCCTCTGGTCCA |
| MLH1 | TAL2328 & TAL2329 | TCTGGCGCCAAAATGTCGttcgtggcaggggttaTTCGGCGGCTGGACGAGA |
| MSH2 | TAL2330 & TAL2331 | TGAGGAGGTTTCGACATGgcggtgcagccgaaggAGACGCTGCAGTTGGAGA |
| MUTYH | TAL2218 & TAL2219 | TCACTGTCGGCGGCCATGacaccgctcgtctcccgcCTGAGTCGTCTGTGGGTA |
| MYC | TAL2398 & TAL2399 | TGCTTAGACGCTGGATTTttttcgggtagtggaaAACCAGGTAAGCACCGAA |
| MYCL1 | TAL2280 & TAL2281 | TCCCGCAGGGAGCGGACAtggactacgactcgtaCCAGCACTATTTCTACGA |
| MYCN | TAL2278 & TAL2279 | TGCCGAGCTGCTCCACgtccaccatgccgggcATGATCTGCAAGAACCCA |
| NBN | TAL2408 & TAL2409 | TGAGGAGCCGGACCGAtgtggaaactgctgccCGCCGCGGGCCCGGCA |
| NCOR1 | TAL2368 & TAL2369 | TCTTTACTGATAATGTCAagttcaggttatcctcCCAACCAAGGAGCATTCA |
| NCOR2 | TAL2284 & TAL2285 | TGGAGGGCCACTGAGCcccgctacccgccccaCAGCCTTTCCTACCCA |
| PDGFRA | TAL2340 & TAL2341 | TTGCGCTCGGGGCGGCCAtgtcggccggcgaggtCGAGCGCCTAGTGTCGGA |
| PDGFRB | TAL2448 & TAL2449 | TCTGCAGGACACCATGCGgcttccgggtgcgatgCCAGCTCTGGCCCTCAAA |
| PHF8 | TAL2246 & TAL2247 | TGAGTACTCCGCCTCTACcccggctgaagcccgcCCCCGCCGCCACCTATTA |
| PMS2 | TAL2418 & TAL2419 | TCGGGTGTTGCATCCATGgagcgagctgagagctcgAGGTGAGCGGGGCTCGCA |
| PTCH1 | TAL2342 & TAL2343 | TGGAACTGCTTAATAGaaacaggcttgtaattGTGAGTCCGCGCTGCA |
| PTEN | TAL2344 & TAL2345 | TCCCAGACATGACAGCCatcatcaaagagatcgTTAGCAGAAACAAAAGGA |
| RARA | TAL2420 & TAL2421 | TGGCATGGCCAGCAACAGcagctcctgcccgacacCTGGGGGCGGGCACCTCA |
| RBBP5 | TAL2370 & TAL2371 | TGCTGGGTGAGAAGGGCtgtggctgcgttttagaGAAGCGTTGGGTACTGGA |
| RECQL4 | TAL2220 & TAL2221 | TGCGGGACGTGCGGGAGCggctgcaggcgtgggaGCGCGCGTTCCGACGGCA |
| RET | TAL2346 & TAL2347 | TGAGTTCTGCCGGCCGCCggctcccgcaggggccaGGGCGAAGTTGGCGCCGA |
| RUNX1 | TAL2452 & TAL2453 | TTCAGGAGGAAGCGATGGcttcagacagcatattTGAGTCATTTCCTTCGTA |
| SDHB | TAL2222 & TAL2223 | TCTCCTTGAGGCGCCGGTtgccggccacaaccctTGGCGGAGCCTGCCTGCA |
| SDHC | TAL2228 & TAL2229 | TGTTGCTGAGGTGACTTCagtgggactgggagttggtGCCTGCGGCCCTCCGGA |
| SDHD | TAL2230 & TAL2231 | TCAGGAACGAGATGGCGGttctctggaggctgagtGCCGTTTGCGGTGCCCTA |
| SETDB1 | TAL2374 & TAL2375 | TGCAGAGGACAAAAGCATgtcttcccttcctgggTGCATTGGTTTGGATGCA |
| SIRT6 | TAL2454 & TAL2455 | TTACGCGGCGGGGCTGTCgccgtacgcggacaagggCAAGTGCGGCCTCCCGGA |
| SMAD2 | TAL2422 & TAL2423 | TTTGGTAAGAACATGTCGtccatcttgccattcacGCCGCCAGTTGTGAAGA |
| SS18 | TAL2442 & TAL2443 | TGGTGACGGCGGCAACATgtctgtggctttcgcggCCCCGAGGCAGCGAGGCA |
| SUZ12 | TAL2376 & TAL2377 | TGGCGCCTCAGAAGCAcggcggtgggggagggGGCGGCTCGGGGCCCA |
| TFE3 | TAL2424 & TAL2425 | TCATGTCTCATGCGGCCGaaccagctcgggatggCGTAGAGGCCAGCGCGGA |
| TP53 | TAL2348 & TAL2472 | TTGCCGTCCCAAGCAATGgatgatttgatgctgtcCCCGGACGATATTGAACA |
| TSC2 | TAL2396 & TAL2397 | TCCTGGTCCACCATGGCcaaaccaacaagcaaagATTCAGGCTTGAAGGAGA |
| VHL | TAL2456 & TAL2457 | TCTGGATCGCGGAGGGAAtgccccggagggcggaGAACTGGGACGAGGCCGA |
| XPA | TAL2458 & TAL2459 | TGGGCCAGAGATGGCGGCggccgacggggctttgCCGGAGGCGGCGGCTTTA |
| XPC | TAL2350 & TAL2351 | TGCCCAGACAAGCAACATggctcggaaacgcgcggccGGCGGGGAGCCGCGGGGA |