Trans-activating crRNA

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tracrRNA at Cas9 (green)

Trans-activating crRNA (tracrRNA) is an RNA that occurs in bacteria as part of the antiviral adaptive immune system CRISPR and is used for the CRISPR / Cas method , CRISPRi and CRISPRa .

properties

The tracrRNA is encoded in trans . It was first described in 2011 in Streptococcus pyogenes . The tracrRNA binds to the crRNA via base pairing and is part of the ribonucleoprotein complex of most Cas proteins (for example Cas9 , Cas12b ). The RNA double strand of crRNA and tracrRNA is converted into the active form via a cut through RNase III and serves the CRISPR / Cas as an adapter for binding the target DNA to be cut. To simplify the use in a CRISPR-based method as the CRISPR / Cas method CRISPRi and CRISPRa is mostly instead of the two RNA (tracrRNA and crRNA) a sgRNA used.

tracrRNA sequences

Type sequence
Streptococcus pyogenes tracrRNA ( Cas9 ) 5'-GUUGGAACCAUUCAAAACAGCAUAGCAAGUUAAAAUAAGGCUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUU-3'
Streptococcus pyogenes tracrDNA ( Cas9 ) 5'-GTTGGAACCATTCAAAACAGCATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTT-3'
Staphylococcus aureus tracrRNA ( Cas9 ) 5'-AUUGUACUUAUACCUAAAAUUACAGAAUCUACUAAAACAAGGCAAAAUGCCGUGUUUAUCUCGUCAACUUGUUGGCGAGAUUUUU-3'
Staphylococcus aureus tracrDNA ( Cas9 ) 5'-ATTGTACTTATACCTAAAATTACAGAATCTACTAAAACAAGGCAAAATGCCGTGTTTATCTCGTCAACTTGTTGGCGAGATTTTT-3'
Streptococcus thermophilus tracrRNA 5'-CUUACACAGUUACUUAAAUCUUGCAGAAGCUACAAAGAUAAGGCUUCAUGCCGAAAUCAACACCCUGUCAUUUUAUGGCAGGGUGUUUU-3'
Streptococcus thermophilus tracrDNA 5'-CTTACACAGTTACTTAAATCTTGCAGAAGCTACAAAGATAAGGCTTCATGCCGAAATCAACACCCTGTCATTTTATGGCAGGGTGTTTT-3'
Corynebacterium diphtheriae tracrRNA 5'-AGUCACUAACUUAAUUAAAUAGAACUGAACCUCAGUAAGCAUUGGCUCGUUUCCAAUGUUGAUUGCUCCGCCGGUGCUCCUUAUUUUUAAGGGCGCCGGCUUUCUU-3'
Corynebacterium diphtheriae tracrDNA 5'-AGTCACTAACTTAATTAAATAGAACTGAACCTCAGTAAGCATTGGCTCGTTTCCAATGTTGATTGCTCCGCCGGTGCTCCTTATTTTTAAGGGCGCCGGCTTTCTT-3'
Alicyclobacillus acidoterrestris tracrRNA ( Cas12b ) 5'-GUCUAGAGGACAGAAUUUUUCAACGGGUGUGCCAAUGGCCACUUUCCAGGUGGCAAAGCCCGUUGAGCUUCUCAAAAAA-3'
Alicyclobacillus acidoterrestris tracrDNA ( Cas12b ) 5'-GTCTAGAGGACAGAATTTTTCAACGGGTGTGCCAATGGCCACTTTCCAGGTGGCAAAGCCCGTTGAGCTTCTCAAAAAA-3'

Individual evidence

  1. ↑ E. Deltcheva, K. Chylinski, CM Sharma, K. Gonzales, Y. Chao, ZA Pirzada, MR Eckert, J. Vogel, E. Charpentier: CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III. In: Nature . Volume 471, number 7340, March 2011, pp. 602-607, doi : 10.1038 / nature09886 , PMID 21455174 , PMC 3070239 (free full text).
  2. ↑ M. Jinek, K. Chylinski, I. Fonfara, M. Hauer, JA Doudna, E. Charpentier: A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. In: Science . Volume 337, number 6096, August 2012, pp. 816-821, doi : 10.1126 / science.1225829 , PMID 22745249 , PMC 6286148 (free full text).
  3. ^ S. Shen, TJ Loh, H. Shen, X. Zheng, H. Shen: CRISPR as a strong gene editing tool. In: BMB reports. Volume 50, number 1, January 2017, pp. 20-24, PMID 27616359 , PMC 5319660 (free full text).
  4. ↑ F. Jiang, JA Doudna: CRISPR-Cas9 Structures and Mechanisms. In: Annual review of biophysics. Volume 46, 05 2017, pp. 505-529, doi : 10.1146 / annurev-biophys-062215-010822 , PMID 28375731 .
  5. ↑ a b c d F. A. Ran, L. Cong, WX Yan, DA Scott, JS Gootenberg, AJ Kriz, B. Zetsche, O. Shalem, X. Wu, KS Makarova, EV Koonin, PA Sharp, F. Zhang: In vivo genome editing using Staphylococcus aureus Cas9. In: Nature . Volume 520, number 7546, April 2015, pp. 186–191, doi : 10.1038 / nature14299 , PMID 25830891 , PMC 4393360 (free full text).
  6. ↑ WO2017222834A1 - Compositions and methods for mitochondrial genome editing - Google Patents. In: patents.google.com. June 10, 2016, accessed January 28, 2019 .
  7. ↑ S. Shmakov, OO Abudayyeh, KS Makarova, YI Wolf, JS Gootenberg, E. Semenova, L. Minakhin, J. Joung, S. Konermann, K. Severinov, F. Zhang, EV Koonin: Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas systems. In: Molecular cell. Volume 60, number 3, November 2015, pp. 385-397, doi : 10.1016 / j.molcel.2015.10.008 , PMID 26593719 , PMC 4660269 (free full text).