I-CreI (m)

Type

Engineered


Target Sequence

-11  -10  -9   -8   -7   -6   -5   -4   -3   -2   -1   1    2    3    4    5    6    7    8    9    10   11  
G A A A A C G T C G T G A G A C A G T T T C

FASTA Sequence



MDTKYNKEFLLYLAGFVDGDGSIIAQIKPNQSYKFKHQLSLTFQVTQKTQRRWFLDKLVDEIGVGYVRDRGSVSD
YILSEIKPLHNFLTQLQPFLKLKQKQANLVLKIIEQLPSAKESPDKFLEVCTWVDQIAALNDSKTRKTTSETVRA
VLDSLSEKKKSSPTGSGSGSKSQAVAHPTDGQRDFGAKGSGSGSGTMNTKYNKEFLLYLAGFVDGDGSIIAQIKP
NQSYKFKHQLSLTFQVTQKTQRRWFLDKLVDEIGVGYVRDRGSVSDYILSEIKPLHNFLTQLQPFLKLKQKQANL
VLKIIEQLPSAKESPDKFLEVCTWVDQIAALNDSKTRKTTSETVRAVLDSLSEKKKSSP

Position Weight Matrix

I-CreI (m)
posACGT
-11 0.25 0.25 0.25 0.25
-10 1.00 0.36 0.28 0.00
-9 1.00 0.20 0.25 0.09
-8 1.00 0.58 0.56 0.30
-7 1.00 1.03 0.00 0.22
-6 0.00 1.00 0.00 1.05
-5 0.69 0.32 1.00 0.61
-4 0.31 0.16 0.00 1.00
-3 0.00 1.00 0.00 0.17
-2 1.21 0.67 1.00 0.68
-1 0.00 0.59 0.52 1.00
1 1.20 0.00 1.00 0.58
2 1.00 0.99 0.79 0.00
3 0.00 0.00 1.00 0.00
4 1.00 0.00 0.29 0.22
5 0.00 1.00 0.28 0.21
6 1.00 0.25 0.81 0.00
7 0.40 0.00 1.00 0.96
8 0.94 0.23 0.27 1.00
9 0.44 0.19 0.02 1.00
10 0.00 0.08 0.32 1.00
11 0.25 0.25 0.25 0.25

Wild-Type Amino Acid Contact Modules

ModuleAmino Acid Contact Modules (?)
-11 → -9:Lys 33, Gln 38
-8 → -6:Gln 26, Lys 28

Key
Experiment
Crystal Structure
Sequence Alignment
* Uncertain


Specificity Changing Mutations (Wild-Type I-CreI)

Amino Acid Mutations (?)                 Specificity Changes                                          Reference
Y33H -10A to -10G, +10T to +10C 1
Y33R -10A to -10G, +10T to +10C 1
Y33C -10A to -10T, +10T to +10A 1
Q26A -6C to -6A, +6A to +6T 2
Q26C -6C to -6G, +6A to +6C 2
Q26C, Y66R -6C to -6G, +6A to +6C 2

1     Seligman LM, Chisholm KM, Chevalier BS, Chadsey MS, Edwards ST, Savage JH, Veillet AL. (2002) Mutations altering the cleavage specificity of a homing endonuclease. Nucleic Acids Research, 30, 3870-3879.
2     Sussman D, Chadsey M, Fauce S, Engel A, Bruett A, Monnat R, Stoddard BL, Seligman LM. (2004) Isolation and characterization of new homing endonuclease specificities at individual target site positions. Journal of Molecular Biology, 342, 31-41.