A recombineering pipeline to make conditional targeting constructs.
Here we describe an optimized recombineering protocol for the creation of conditional alleles. Recombineering is a DNA engineering methodology based on homologous recombination in Escherichia coli. It presents a number of advantages over conventional engineering methods, including the ability to place target sites for site-specific recombinases at exactly chosen positions. Consequently, complicated designs, such as conditional alleles, can be rapidly achieved. We have applied these advantages to establish a pipeline method for the fluent generation of conditional alleles which employs Cre, Flp, and Dre recombinase target sites.
Fu
Jun
J
Genomics, BioInnovationsZentrum, Technische Universitaet Dresden, Dresden, Germany.
Teucher
Madeleine
M
Anastassiadis
Konstantinos
K
Skarnes
William
W
Stewart
A Francis
AF
D000818:Animals; D004254:DNA Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods*; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D011995:Recombination, Genetic / Q000235:genetics*
C045073:Cre recombinase; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase; D019426:Integrases; C077570:Site-specific recombinase