High resolution mapping and positional cloning of ENU-induced mutations in the Rw region of mouse chromosome 5.
BACKGROUND
Forward genetic screens in mice provide an unbiased means to identify genes and other functional genetic elements in the genome. Previously, a large scale ENU mutagenesis screen was conducted to query the functional content of a ~50 Mb region of the mouse genome on proximal Chr 5. The majority of phenotypic mutants recovered were embryonic lethals.


RESULTS
We report the high resolution genetic mapping, complementation analyses, and positional cloning of mutations in the target region. The collection of identified alleles include several with known or presumed functions for which no mutant models have been reported (Tbc1d14, Nol14, Tyms, Cad, Fbxl5, Haus3), and mutations in genes we or others previously reported (Tapt1, Rest, Ugdh, Paxip1, Hmx1, Otoe, Nsun7). We also confirmed the causative nature of a homeotic mutation with a targeted allele, mapped a lethal mutation to a large gene desert, and localized a spermiogenesis mutation to a region in which no annotated genes have coding mutations. The mutation in Tbc1d14 provides the first implication of a critical developmental role for RAB-GAP-mediated protein transport in early embryogenesis.


CONCLUSIONS
This collection of alleles contributes to the goal of assigning biological functions to all known genes, as well as identifying novel functional elements that would be missed by reverse genetic approaches.
Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853, USA.
Yung-Hao
YH
Ching
Robert J
RJ
Munroe
Jennifer L
JL
Moran
Anna K
AK
Barker
Evan
E
Mauceli
Tim
T
Fennell
Frederica
F
Dipalma
Kerstin
K
Lindblad-Toh
Lindsay M
LM
Abcunas
Joyanna F
JF
Gilmour
Tanya P
TP
Harris
Susan L
SL
Kloet
Yunhai
Y
Luo
John L
JL
McElwee
Weipeng
W
Mu
Hyo K
HK
Park
David L
DL
Rogal
Kerry J
KJ
Schimenti
Lishuang
L
Shen
Mami
M
Shindo
James Y
JY
Shou
Erin K
EK
Stenson
Patrick J
PJ
Stover
John C
JC
Schimenti
D000818:Animals; D002874:Chromosome Mapping; D002875:Chromosomes / Q000187:drug effects*; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D047108:Embryonic Development / Q000235:genetics*; D005038:Ethylnitrosourea / Q000633:toxicity; D005804:Genes, Lethal; D005816:Genetic Complementation Test; D008297:Male; D051379:Mice / Q000235:genetics*; D008810:Mice, Inbred C57BL; D009154:Mutation; D017384:Sequence Deletion; D013091:Spermatogenesis / Q000235:genetics
D005038:Ethylnitrosourea