MARCM

Mosaic Analysis with a Repressible Cell Marker system

This collection consists of about 2,200 strains possessing the autosomal insertions of a new piggyBac vector (PBac{SAstopDsRed}), made by the laboratory of Professor L. Luo at Stanford University. The strains have following properties:

  • All strains have a insertion inside independent transcriptional units.
  • All insertions are on FRT-bearing chromosomes.
  • The piggyBac vector have splicing acceptor and a termination of translation in all three frames, thus improving mutagenicity.
  • The marker on the vector is a DsRed protein, enabling easier viewing of mosaic clones labeled with GFP.

>>> Stock list

Insertions on the second chromosome:

Genotype:
y* w*; PBac{SAstopDsRed}LL***** P{neoFRT}40A P{FRT(whs)}G13 cn1 bw1/CyO, S* bw1

In some stocks, there are floating third chromosome,
P{FRT(whs)}2A P{neoFRT}82B P{y+t7.7 ry+t7.2=Car20y}96E,
TM6B, Tb1,
or both.

IMPORTANT NOTES:

1) The combination of cn1 bw1/CyO, S* bw1 results in white eyes *1 regardless of w+ transgenes - therefore all flies should have white eyes when in stock - cross out to see red eye.
2) Third chromosome was never actively selected against, so y+ flies may exist.
3) No FRT on mutator vector - thus NOT compatible with Harvard collection to create deletions.

Thanks to Oren Schuldiner (Weizmann Institute of Science)

Insertions on the third chromosome:

Genotype:
y* w*; PBac{SAstopDsRed}LL***** P{FRT(whs)}2A P{neoFRT}82B P{y+t7.7 ry+t7.2=Car20y}96E/TM6B, Tb1

In some stocks, there are floating second chromosome,
P{neoFRT}40A P{FRT(whs)}G13 cn1 bw1,
CyO, S[*] bw1,
or both
(this case white-eyed flies will be found).

IMPORTANT NOTES:

1) Because second chromosome was never actively selected against, a small percentage of homozygous cn bw flies resulting in white eyes can be present in the population.
2) All third chromosome flies should be yellow+ due to the insertion at 96E.
3) A small percentage of early insertions (lower LL number) were balanced with TM3, Sb1 instead of TM6, Tb1.
4) No FRT on mutator vector - thus NOT compatible with Harvard collection to create deletions.

Thanks to Oren Schuldiner (Weizmann Institute of Science)

Mutator Vector

FlyBase: PBac{SAstopDsRed}

GenBank Accesstion Number: EU257621

Schematic map of PBac{SAstopDsRed} from Oren Schuldiner (Weizmann Institute of Science)
pB_vector_DsRed_SA_StopB_W-s.jpg

Reference

Oren Schuldiner, Daniela Berdnik, Jonathan Ma Levy, Joy S. Wu, David Luginbuhl, Allison Camille Gontang and Liqun Luo. piggyBac-Based Mosaic Screen Identifies a Postmitotic Function for Cohesin in Regulating Developmental Axon Pruning. Developmental Cell Volume 14, Issue 2, 12 February 2008, Pages 227-238 doi:10.1016/j.devcel.2007.11.001


*1 cn bw is white eyes! CyO has cn2 as a recessive marker. Genotype of CyO is In(2LR)O, Cy1 dplv1 pr1 cn2, which is not usually described in the genotype of the strain. Thus, CyO, S* bw1 has markers as S* Cy1 dplv1 pr1 cn2 bw1. See also FBba0000025

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Last-modified: 10 Apr 2010 (Sat) 11:19:38 (1620d)